Method for the detection of type II diabetes related gene transcripts in blood

ABSTRACT

The present invention is directed to detection and measurement of gene transcripts and their equivalent nucleic acid products in blood. Specifically provided is analysis performed on a drop of blood for detecting, diagnosing and monitoring diseases using gene-specific and/or tissue-specific primers. The present invention also describes methods by which delineation of the sequence and/or quantitation of the expression levels of disease-specific genes allows for an immediate and accurate diagnostic/prognostic test for disease or to assess the effect of a particular treatment regimen.

RELATED APPLICATIONS

[0001] This application is a Divisional of Application of: Choong-ChinLiew, Filed: Mar. 12, 2004, Ser. No.: ______, Entitled: A Method for theDetection of Coronary Artery Disease Related Gene Transcripts in Blood,Our Reference No.: 4231/2055B, which a continuation in part ofapplication Ser. No. 10/601,518, filed on Jun. 20, 2003, which is acontinuation-in-part of application Ser. No. 10/085,783, filed on Feb.28, 2002, which claims the benefit of U.S. Provisional Application No.60/271,955, filed on Feb. 28, 2001, U.S. Provisional Application No.60/275,017 filed Mar. 12, 2001, and U.S. Provisional Application No.60/305,340; filed July 132001, and is also a continuation-in-part ofapplication Ser. No. 10/268,730 filed on Oct. 9, 2002, which is acontinuation of U.S. application Ser. No. 09/477,148 filed Jan. 4, 2000,now abandoned, which claims the benefit of U.S. Provisional ApplicationNo. 60/115,125 filed on Jan. 6, 1999. Each of these applications isincorporated herein by reference in their entirety, including figuresand drawings.

[0002] Tables

[0003] This application includes a compact disc in duplicate (2 compactdiscs: Tables—Copy 1 and Tables—Copy 2), which are hereby incorporatedby reference in their entirety. Each compact disc is identical andcontains the following files (corresponding to Tables 2-4): TABLEDESCRIPTION SIZE CREATED Text File Name 1 2 multi-gene comparison371,563 Mar. 25, 2004 TABLE2.TXT 2 3A GLF 8 - hypertension 138,940 Mar.28, 2004 TABLE3A.TXT 3 3AA GLF 29 - asthma 36,121 Mar. 27, 2004TABLE3AA.TXT 4 3AB multi OA 29,898 Mar. 27, 2004 TABLE3AB.TXT 5 3AC GLMDS vs. schizo 114,078 Mar. 27, 2004 TABLE3AC.TXT 6 3AD steroiddifferential 64,646 Mar. 27, 2004 TABLE3AD.TXT 7 3B GLF 9 - obesity147,421 Mar. 25, 2004 TABLE3B.TXT 8 3C GLF 10 - allergies 95,700 Mar.25, 2004 TABLE3C.TXT 9 3D GLF 11 - steroids 93,808 Mar. 25, 2004TABLE3D.TXT 10 3E GLF 12 - hypertension 314,854 Mar. 25, 2004TABLE3E.TXT 11 3F GLF 13 - obesity 181,310 Mar. 25, 2004 TABLE3F.TXT 123G GLF 14 - diabetes 146,212 Mar. 26, 2004 TABLE3G.TXT 13 3H GLF 15 -hyperlipidemia 165,909 Mar. 26, 2004 TABLE3H.TXT 14 3I GLF 16 - lung92,936 Mar. 25, 2004 TABLE3I.TXT 15 3J GLF 17 - bladder 1,143,423 Mar.26, 2004 TABLE3J.TXT 16 3K GLF 18 - bladder 953,119 Mar. 26, 2004TABLE3K.TXT 17 3L GLF 19 - Coronary Art Dis. 246,178 Mar. 26, 2004TABLE3L.TXT 18 3M GLF 20 - rheumarth 329,672 Mar. 26, 2004 TABLE3M.TXT19 3N GLF 21 - depression 153,108 Mar. 26, 2004 TABLE3N.TXT 20 3O GLF22 - rheumarth 49,043 Mar. 26, 2004 TABLE3O.TXT 21 3P GLF hypertension577 only 84,945 Mar. 26, 2004 TABLE3P.TXT 22 3Q GLF OA hypertensionshared 33,081 Mar. 26, 2004 TABLE3Q.TXT 23 3R GL obesity 519 79,544 Mar.26, 2004 TABLE3R.TXT 24 3S GL obesity shared 152 24,583 Mar. 26, 2004TABLE3S.TXT 25 3T GL allergy specific 39,547 Mar. 25, 2004 TABLE3T.TXT26 3U GL allergy OA shared 241 35,603 Mar. 25, 2004 TABLE3U.TXT 27 3V GLsteroid 362 54,954 Mar. 26, 2004 TABLE3V.TXT 28 3W GL OA steroid shared31,459 Mar. 27, 2004 TABLE3W.TXT 29 3X GLF 26 - liver cancer 435,093Mar. 27, 2004 TABLE3X.TXT 30 3Y GLF 27 - schizophrenia 578,949 Mar. 26,2004 TABLE3Y.TXT 31 3Z GLF 28 - chagas 202,477 Mar. 28, 2004 TABLE3Z.TXT32 4 sequence listing 114,765 Mar. 11, 2004 TABLE4.TXT

BACKGROUND

[0004] The blood is a vital part of the human circulatory system for thehuman body. Numerous cell types make up the blood tissue includingmonocytes, leukocytes, lymphocytes and erythrocytes. Although many bloodcell types have been described, there are likely many as yetundiscovered cell types in the human blood. Some of these undiscoveredcells may exist transiently, such as those derived from tissues andorgans that are constantly interacting with the circulating blood inhealth and disease. Thus, the blood can provide an immediate picture ofwhat is happening in the human body at any given time.

[0005] The turnover of cells in the hematopoietic system is enormous. Itwas reported that over one trillion cells, including 200 billionerythrocytes and 70 billion neutrophilic leukocytes, turn over each dayin the human body (Ogawa 1993). As a consequence of continuousinteractions between the blood and the body, genetic changes that occurwithin the cells or tissues of the body will trigger specific changes ingene expression within blood. It is the goal of the present inventionthat these genetic alterations be harnessed for diagnostic andprognostic purposes, which may lead to the development of therapeuticsfor ameliorating disease.

[0006] For example, isoformic myosin heavy chain genes are known to begenerally expressed in cardiac muscle tissue. In the rodent, the βMyHCgene is only highly expressed in the fetus and in diseased states suchas overt cardiac hypertrophy, heart failure and diabetes; the αMyHC geneis highly expressed shortly after birth and continues to be expressed inthe adult heart. In the human, however, βMyHC is highly expressed in theventricles from the fetal stage through adulthood. This highly expressedβMyHC, which harbours several mutations, has been demonstrated to beinvolved in familial hypertrophic cardiomyopathy (Geisterfer-Lowrance etal. 1990). It was reported that mutations of βMyHC can be detected byPCR using blood lymphocyte DNA (Ferrie et al., 1992). Most recently, itwas also demonstrated that mutations of the myosin-binding protein C infamilial hypertrophic cardiomyopathy can be detected in the DNAextracted from lymphocytes (Niimura et al., 1998).

[0007] Similarly, APP and APC, which are known to be tissue specific andpredominantly expressed in the brain and intestinal tract, are alsodetectable in the transcripts of blood. These cell- or tissue-specifictranscripts are not detectable by Northern blot analysis. However, thelow number of transcript copies can be detected by RT-PCR analysis.These findings strongly demonstrate that genes preferentially expressedin specific tissues can be detected by a highly sensitive RT-PCR assay.In recent years, evidence has been obtained to indicate that expressionof cell or tissue-restricted genes can be detected in the certainperipheral nucleated blood cells of patients with metastatictransitional cell carcinoma (Yuasa et al. 1998) and patients withprostate cancer (Gala et al. 1998).

[0008] In the prior art, there is a need for large samples and/or costlyand time-consuming separation of cell types within the blood (Kimoto(1998) and Chelly et al. (1989; 1988)). The prior art, however, isdeficient in non-invasive methods of screening for tissue-specificdiseases. The present invention fulfills this long-standing need anddesire in the art.

SUMMARY OF THE INVENTION

[0009] The present invention relates generally to the molecular biologyof human diseases. More specifically, the present invention relates to aprocess using the genetic information contained in human peripheralwhole blood for the diagnosis, prognosis and monitoring of genetic andinfectious disease in the human body.

[0010] This present invention discloses a process of using the geneticinformation contained in human peripheral whole blood in the diagnosis,prognosis and monitoring of genetic and infectious disease in the humanbody. The process described herein requires a simple blood sample andis, therefore, non-invasive compared to conventional practices used todetect tissue specific disease, such as biopsies.

[0011] The invention is based on the discovery that gene expression inthe blood is reflective of body state and, as such, the resultantdisruption of homeostasis under conditions of disease can be detectedthrough analysis of transcripts differentially expressed in the bloodalone. Thus, the identification of several key transcripts or geneticmarkers in blood will provide information about the genetic state of thecells, tissues, organ systems of the human body in health and disease.

[0012] The present invention demonstrates that a simple drop of bloodmay be used to determine the quantitative expression of various mRNAsthat reflect the health/disease state of the subject through the use ofRT-PCR analysis. This entire process takes about three hours or less.The single drop of blood may also be used for multiple RT-PCR analyses.It is believed that the present finding can potentially revolutionizethe way that diseases are detected, diagnosed and monitored because itprovides a non-invasive, simple, highly sensitive and quick screeningfor tissue-specific transcripts. The transcripts detected in whole bloodhave potential as prognostic or diagnostic markers of disease, as theyreflect disturbances in homeostasis in the human body. Delineation ofthe sequences and/or quantitation of the expression levels of thesemarker genes by RT-PCR will allow for an immediate and accuratediagnostic/prognostic test for disease or to assess the efficacy andmonitor a particular therapeutic.

[0013] One object of the present invention is to provide a non-invasivemethod for the diagnosis, prognosis and monitoring of genetic andinfectious disease in humans and animals.

[0014] In one embodiment of the present invention, there is provided amethod for detecting expression of a gene in blood from a subject,comprising the steps of: a) quantifying RNA from a subject blood sample;and b) detecting expression of the gene in the quantified RNA, whereinthe expression of the gene in quantified RNA indicates the expression ofthe gene in the subject blood. An example of the quantifying method isby mass spectrometry.

[0015] In another embodiment of the present invention, there is provideda method for detecting expression of one or more genes in blood from asubject, comprising the steps of: a) obtaining a subject blood sample;b) extracting RNA from the blood sample; c) amplifying the RNA; d)generating expressed sequence tags (ESTs) from the amplified RNAproduct; and e) detecting expression of the genes in the ESTs, whereinthe expression of the genes in the ESTs indicates the expression of thegenes in the subject blood. Preferably, the subject is a fetus, anembryo, a child, an adult or a non-human animal. The genes arenon-cancer-associated and tissue-specific genes. Still preferably, theamplification is performed by RT-PCR using random sequence primers orgene-specific primers.

[0016] In still another embodiment of the present invention, there isprovided a method for detecting expression of one or more genes in bloodfrom a subject, comprising the steps of: a) obtaining a subject bloodsample; b) extracting DNA fragments from the blood sample; c) amplifyingthe DNA fragments; and d) detecting expression of the genes in theamplified DNA product, wherein the expression of the genes in theamplified DNA product indicates the expression of the genes in thesubject blood.

[0017] In yet another embodiment of the present invention, there isprovided a method for monitoring a course of a therapeutic treatment inan individual, comprising the steps of: a) obtaining a blood sample fromthe individual; b) extracting RNA from the blood sample; c) amplifyingthe RNA; d) generating expressed sequence tags (ESTs) from the amplifiedRNA product; e) detecting expression of genes in the ESTs, wherein theexpression of the genes is associated with the effect of the therapeutictreatment; and f) repeating steps a)-e), wherein the course of thetherapeutic treatment is monitored by detecting the change of expressionof the genes in the ESTs. Such a method may also be used for monitoringthe onset of overt symptoms of a disease, wherein the expression of thegenes is associated with the onset of the symptoms. Preferably, theamplification is performed by RT-PCR, and the change of the expressionof the genes in the ESTs is monitored by sequencing the ESTs andcomparing the resulting sequences at various time points; or byperforming single nucleotide polymorphism analysis and detecting thevariation of a single nucleotide in the ESTs at various time points.

[0018] In still yet another embodiment of the present invention, thereis provided a method for diagnosing a disease in a test subject,comprising the steps of: a) generating a cDNA library for the diseasefrom a whole blood sample from a normal subject; b) generating expressedsequence tag (EST) profile from the normal subject cDNA library; c)generating a cDNA library for the disease from a whole blood sample froma test subject; d) generating EST profile from the test subject cDNAlibrary; and e) comparing the test subject EST profile to the normalsubject EST profile, wherein if the test subject EST profile differsfrom the normal subject EST profile, the test subject might be diagnosedwith the disease.

[0019] In still yet another embodiment of the present invention, thereis provided a kit for diagnosing, prognosing or predicting a disease,comprising: a) gene-specific primers; wherein the primers are designedin such a way that their sequences contain the opposing ends of twoadjacent exons for the specific gene with the intron sequence excluded;and b) a carrier, wherein the carrier immobilizes the primer(s).Preferably, the gene-specific primers are selected from the groupconsisting of insulin-specific primers, atrial natriureticfactor-specific primers, zinc finger protein gene-specific primers,beta-myosin heavy chain gene-specific primers, amyloid precursor proteingene-specific primers, and adenomatous polyposis-coli proteingene-specific primers. Further preferably, the gene-specific primers areselected from the group consisting of SEQ ID Nos. 1 and 2; and SEQ IDNos. 5 and 6. Such a kit may be applied to a test subject whole bloodsample to diagnose, prognose or predict a disease by detecting thequantitative expression levels of specific genes associated with thedisease in the test subject and then comparing to the levels of samegenes expressed in a normal subject. Such a kit may also be used formonitoring a course of therapeutic treatment or monitoring the onset ofovert symptoms of a disease.

[0020] In yet another embodiment of the present invention, there isprovided a kit for diagnosing, prognosing or predicting a disease,comprising: a) probes derived from a whole blood sample for a specificdisease; and b) a carrier, wherein the carrier immobilizes the probes.Such a kit may be applied to a test subject whole blood sample todiagnose, prognose or predict a disease by detecting the quantitativeexpression levels of specific genes associated with the disease in thetest subject and then comparing to the levels of same genes expressed ina normal subject. Such a kit may also be used for monitoring a course oftherapeutic treatment or monitoring the onset of overt symptoms of adisease.

[0021] Furthermore, the present invention provides a cDNA libraryspecific for a disease, wherein the cDNA library is generated from wholeblood samples.

[0022] In one embodiment of the present invention, there is a method ofidentifying one or more genetic markers for a disease, wherein each ofsaid one or more genetic markers corresponds to a gene transcript,comprising the steps of: a) determining the level of one or more genetranscripts expressed in blood obtained from one or more individualshaving a disease, wherein each of said one or more transcripts isexpressed by a gene that is a candidate marker for disease; and b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals not having adisease, wherein those compared transcripts which display differinglevels in the comparison of step b) are identified as being geneticmarkers for a disease.

[0023] In another embodiment of the present invention, there is a methodof identifying one or more genetic markers for a disease, wherein eachof said one or more genetic markers corresponds to a gene transcript,comprising the steps of: a) determining the level of one or more genetranscripts expressed in blood obtained from one or more individualshaving a disease, wherein each of said one or more transcripts isexpressed by a gene that is a candidate marker for a disease; and b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals having adisease, wherein those compared transcripts which display the samelevels in the comparison of step b) are identified as being geneticmarkers for a disease.

[0024] In another embodiment of the present invention, there is a methodof identifying one or more genetic markers of a stage of a diseaseprogression or regression, wherein each of said one or more geneticmarkers corresponds to a gene transcript, comprising the steps of: a)determining the level of one or more gene transcripts expressed in bloodobtained from one or more individuals having a stage of a disease,wherein said one or more individuals are at the same progressive orregressive stage of a disease, and wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordetermining the stage of progression or regression of a disease, and; b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals who are at aprogressive or regressive stage of a disease distinct from that of saidone or more individuals of step a), wherein those compared transcriptswhich display differing levels in the comparison of step b) areidentified as being genetic markers for the stage of progression orregression of a disease.

[0025] In another embodiment of the present invention, there is a methodof identifying one or more genetic markers of a stage of a diseaseprogression or regression, wherein each of said one or more geneticmarkers corresponds to a gene transcript, comprising the steps of: a)determining the level of one or more gene transcripts expressed in bloodobtained from one or more individuals having a stage of a disease,wherein said one or more individuals are at the same progressive orregressive stage of a disease, and wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordetermining the stage of progression or regression of a disease, and b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals who are at aprogressive or regressive stage of a disease identical to that of saidone or more individuals of step a), wherein those compared transcriptswhich display the same levels in the comparison of step b) areidentified as being genetic markers for the stage of progression orregression of a disease.

[0026] Further embodiments of the methods described in the previous fourparagraphs include the embodiments wherein each of said one or moremarkers identifies one or more transcripts of one or more non immuneresponse genes, wherein each of said one or more markers identifies atranscript of a gene expressed by non-blood tissue, wherein each of saidone or more markers identifies a transcript of a gene expressed bynon-lymphoid tissue, wherein said one or more markers identifies asequence selected from the sequences listed in any one of Table 3A-Z andTables 3AA, 3AB, 3AC and 3AD, wherein said one or more markersidentifies the sequence of one or more of the sequences selected fromthe group consisting of ANF, ZFP and βMyHC, wherein said blood comprisesa blood sample obtained from said one or more individuals, wherein saidblood sample consists of whole blood, wherein said blood sample consistsof a drop of blood, and wherein said blood sample consists of blood thathas been lysed.

[0027] In another embodiment of the present invention, there is a methodof diagnosing or prognosing a disease in an individual, comprising thesteps of: a) determining the level of one or more gene transcripts inblood obtained from said individual suspected of having a disease, andb) comparing the level of each of said one or more gene transcripts insaid blood according to step a) with the level of each of said one ormore gene transcripts in blood from one or more individuals not having adisease, wherein detecting a difference in the levels of each of saidone or more gene transcripts in the comparison of step b) is indicativeof a disease in the individual of step a).

[0028] In another embodiment of the present invention, there is a methodof diagnosing or prognosing a disease in an individual, comprising thesteps of: a) determining the level of one or more gene transcripts inblood obtained from said individual suspected of having a disease, andb) comparing the level of each of said one or more gene transcripts insaid blood according to step a) with the level of each of said one ormore gene transcripts in blood from one or more individuals having adisease, wherein detecting the same levels of each of said one or moregene transcripts in the comparison of step b) is indicative of a diseasein the individual of step a).

[0029] In another embodiment of the present invention, there is a methodof determining a stage of disease progression or regression in anindividual having a disease, comprising the steps of: a) determining thelevel of one or more gene transcripts in blood obtained from saidindividual having a disease, and b) comparing the level of each if saidone or more gene transcripts in said blood according to step a) with thelevel of each of said one or more gene transcripts in blood obtainedfrom one or more individuals who each have been diagnosed as being atthe same progressive or regressive stage of a disease, wherein thecomparison from step b) allows the determination of the stage of adisease progression or regression in an individual.

[0030] In another embodiment of the present invention, there is a methodof diagnosing or prognosing osteoarthritis in an individual, comprisingthe steps of: a) determining the level of one or more gene transcriptsexpressed in blood obtained from said individual, wherein said one ormore gene transcripts correspond to said one or more markers of claim 1and claim 2, and b) comparing the level of each of said one or more genetranscripts in said blood according to step a) with the level of each ofsaid one or more gene transcripts in blood from one or more individualshaving osteoarthritis, c) comparing the level of each of said one ormore gene transcripts in said blood according to step a) with the levelof each of said one or more gene transcripts in blood from one or moreindividuals not having osteoarthritis, d) determining whether the levelof said one or more gene transcripts of step a) classify with the levelsof said transcripts in step b) as compared with the levels of saidtranscripts in step c) wherein said determination is indicative of saidindividual of step a) having osteoarthritis.

[0031] In another embodiment of the present invention, there is a methodof determining a stage of disease progression or regression in anindividual having osteoarthritis, comprising the steps of: a)determining the level of one or more gene transcripts expressed in bloodobtained from said individual having said stage of osteoarthritis,wherein said one or more gene transcripts correspond to the markers ofclaim 3 and claim 4, and b) comparing the level of each of said one ormore gene transcripts in said blood according to step a) with the levelof each of said one or more gene transcripts in blood from one or moreindividuals having said stage of osteoarthritis, c) comparing the levelof each of said one or more gene transcripts in said blood according tostep a) with the level of each of said one or more gene transcripts inblood from one or more individuals not having said stage ofosteoarthritis, d) determining whether the level of said one or moregene transcripts of step a) classify with the levels of said transcriptsin step b) as compared with levels of said transcripts in step c),wherein said determination is indicative of said individual of step a)having said stage of osteoarthritis.

[0032] Further embodiments of the methods described in the previous tenparagraphs include embodiments comprising a further step of isolatingRNA from said blood samples, and embodiments comprising determining thelevel of each of said one or more gene transcripts comprisingquantitative RT-PCR (QRT-PCR), wherein said one or more transcripts arefrom step a) and/or step b) of said methods. Further embodiments ofthese methods include embodiments wherein said QRT-PCR comprises primerswhich hybridize to one or more transcripts or the complement thereof,wherein said one or more transcripts are from step a) and/or step b) ofsaid methods, embodiments wherein said primers are 15-25 nucleotides inlength, and embodiments wherein said primers hybridize to one or more ofthe sequences of any one of Table 3A-Z and Tables 3AA, 3AB, 3AC and 3AD,or the complement thereof. Further embodiments of the methods describedin the previous eight paragraphs include embodiments wherein the step ofdetermining the level of each of said one or more gene transcriptscomprises hybridizing a first plurality of isolated nucleic acidmolecules that correspond to said one or more transcripts to an arraycomprising a second plurality of isolated nucleic acid molecules,wherein in one embodiment said first plurality of isolated nucleic acidmolecules comprises RNA, DNA, cDNA, PCR products or ESTs, wherein in oneembodiment said array comprises a plurality of isolated nucleic acidmolecules comprising RNA, DNA, cDNA, PCR products or ESTs, wherein inone embodiment said array comprises two or more of the genetic markersof said methods, wherein in one embodiment said array comprises aplurality of nucleic acid molecules that correspond to genes of thehuman genome.

[0033] In another embodiment of the present invention, there is aplurality of nucleic acid molecules that correspond to two or moresequences from each of any one of Table 3A-Z and Tables 3AA, 3AB, 3ACand 3AD.

[0034] In another embodiment of the present invention, there is an arraywhich comprises a plurality of nucleic acid molecules that correspond totwo or more sequences from each of any one of Table 3A-Z and Tables 3AA,3AB, 3AC and 3AD.

[0035] In another embodiment of the present invention, there is a kitfor diagnosing or prognosing a disease comprising: a) two gene-specificpriming means designed to produce double stranded DNA complementary to agene selected from the group consisting of Table 3L; wherein said firstpriming means contains a sequence which can hybridize to RNA, cDNA or anEST complementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity c) an enzyme withthermostable DNA polymerase activity and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject

[0036] In another embodiment of the present invention, there is a kitfor monitoring a course of therapeutic treatment of a disease,comprising a) two gene-specific priming means designed to produce doublestranded DNA complementary to a gene selected group consisting of anyone of Table 3A-Z and Tables 3AA, 3AB, 3AC and 3AD; wherein said firstpriming means contains a sequence which can hybridize to RNA, cDNA or anEST complementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity c) an enzyme withthermostable DNA polymerase activity and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject.

[0037] In another embodiment of the present invention, there is a kitfor monitoring progression or regression of a disease, comprising: a)two gene-specific priming means designed to produce double stranded DNAcomplementary to a gene selected group consisting of any one of Table3A-Z and Tables 3AA, 3AB, 3AC and 3AD; wherein said first priming meanscontains a sequence which can hybridize to RNA, cDNA or an ESTcomplementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity c) an enzyme withthermostable DNA polymerase activity and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject.

[0038] In another embodiment of the present invention, there is aplurality of nucleic acid molecules that identify or correspond to twoor more sequences from any one of Table 3A-Z and Tables 3AA, 3AB, 3ACand 3AD.

[0039] Other and further aspects, features, and advantages of thepresent invention will be apparent from the following description of thepresently preferred embodiments of the invention. These embodiments aregiven for the purpose of disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The above-recited features, advantages and objects of theinvention, as well as others which will become clear, are attained andcan be understood in detail, more particular descriptions of theinvention briefly summarized above may be had by reference to certainembodiments thereof which are illustrated in the appended drawings.These drawings form a part of the specification. It is to be noted,however, that the appended drawings illustrate preferred embodiments ofthe invention and therefore are not to be considered limiting in theirscope.

[0041]FIG. 1 shows the following RNA samples prepared from human blood;FIG. 1A: Lane 1, Molecular weight marker; Lane 2, RT-PCR on APP gene;Lane 3, PCR on APP gene; Lane 4, RT-PCR on APC gene; Lane 5, PCR on APCgene; FIG. 1B: Lanes 1 and 2, RT-PCR and PCR of βMyHC, respectively;Lanes 3 and 4, RT-PCR of βMyHC from RNA prepared from human fetal andhuman adult heart, respectively; Lane 5, Molecular weight marker.

[0042]FIG. 2 shows quantitative RT-PCR analysis performed on RNA samplesextracted from a drop of blood. Forward primer(5′-GCCCTCTGGGGACCTGAC-3′, SEQ ID No. 1) of exon 1 and reverse primer(5′-CCCACCTGCAGGTCCTCT-3″, SEQ ID No. 2) of exons 1 and 2 of insulingene. Blood samples of 4 normal subjects were assayed. Lanes 1, 3, 5 and7 represent overnight “fasting” blood sample and lanes 2, 4, 6 and 8represent “non-fasting” samples.

[0043]FIG. 3 shows quantitative RT-PCR analysis performed on RNA samplesextracted from a drop of blood. Lanes 1 and 2 represent normal healthyperson and lane 3 represents late-onset diabetes (Type II) and lane 4represents asymptomatic diabetes.

[0044]FIG. 4 shows multiple RT-PCR assay in a drop of blood. Primerswere derived from insulin gene (INS), zinc-finger protein gene (ZFP) andhouse-keeping gene (GADH). Lane 1 represents normal person. Lane 2represents late-onset diabetes and lane 3 represents asymptomaticdiabetes.

[0045]FIG. 5 shows standardized levels of insulin gene (FIG. 5A) and ZFPgene (FIG. 5B) expressed in a drop of blood. The first three subjectswere normal, second two subjects showed normal glucose tolerance, andthe last subject had late onset diabetes type II. FIG. 5C showsstandardized levels of insulin gene expressed in each fractionated cellfrom whole blood.

[0046]FIG. 6 shows the differential screening of human blood cell cDNAlibrary with different cDNA probes of heart and brain tissue. FIG. 6Ashows blood cell cDNA probes vs. adult heart cDNA probes. FIG. 6B showsblood cell cDNA probes vs. human brain cDNA probes.

[0047]FIG. 7 graphically shows the 1,800 unique genes in human blood andin the human fetal heart grouped into seven cellular functions.

[0048]FIG. 8 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having both osteoarthritisand hypertension as compared with gene expression profiles from normalindividuals.

[0049]FIG. 9 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingboth osteoarthritis and who were obese as described herein as comparedwith gene expression profiles from normal individuals

[0050]FIG. 10 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingboth osteoarthritis and allergies as described herein as compared withgene expression profiles from normal individuals.

[0051]FIG. 11 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having osteoarthritis and whowere subject to systemic steroids as described herein as compared withgene expression profiles from normal individuals.

[0052]FIG. 12 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having hypertension ascompared with gene expression profiles from samples of bothnon-hypertensive and normal individuals.

[0053]FIG. 13 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as obeseas described herein as compared with gene expression profiles fromnormal and non-obese individuals.

[0054]FIG. 14 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingtype 2 diabetes as described herein as compared with gene expressionprofiles from normal and non-type 2 diabetes individuals.

[0055]FIG. 15 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havinghyperlipidemia as described herein as compared with gene expressionprofiles from normal and non-hyperlipidemia patients.

[0056]FIG. 16 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havinglung disease as described herein as compared with gene expressionprofiles from normal and non lung disease individuals.

[0057]FIG. 17 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingbladder cancer as described herein as compared with gene expressionprofiles from non bladder cancer individuals.

[0058]FIG. 18 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingadvanced stage bladder cancer or early stage bladder cancer as describedherein as compared with gene expression profiles from non bladder cancerindividuals.

[0059]FIG. 19 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingcoronary artery disease (CAD) as described herein as compared with geneexpression profiles from non-coronary artery disease individuals.

[0060]FIG. 20 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingrheumatoid arthritis as described herein as compared with geneexpression profiles from non-rheumatoid arthritis individuals.

[0061]FIG. 21 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingdepression as described herein as compared with gene expression profilesfrom non-depression individuals.

[0062]FIG. 22 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingvarious stages of osteoarthritis as described herein as compared withgene expression profiles from normal individuals.

[0063]FIG. 23 shows RT-PCR of overexpressed genes in CAD peripheralblood cells identified using microarray experiments, including PBP, PF4and F13A.

[0064]FIG. 24 shows the “Blood Chip”, a cDNA microarray slide with10,368 PCR products derived from peripheral blood cell cDNA libraries.Colors represent hybridization to probes labelled with Cy3 (green) orCy5 (red). Yellow spots indicate common hybridization between bothprobes. In slide A, normal blood cell RNA samples were labelled with Cy3and CAD blood cell RNA samples were labelled with Cy5. In slide B, Cy3and Cy5 were switched to label the RNA samples. (Cluster analysisrevealed distinct gene expression profiles for normal and CAD samples.)

[0065]FIG. 25 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingliver cancer as described herein as compared with gene expressionprofiles from normal individuals.

[0066]FIG. 26 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingschizophrenia as described herein as compared with gene expressionprofiles from normal individuals.

[0067]FIG. 27 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingsymptomatic or asymptomatic chagas disease as described herein ascompared with gene expression profiles from normal individuals.

[0068]FIG. 28 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingasthma and OA as compared with individuals having just OA.

[0069]FIG. 29 shows a venn diagram illustrating a summary of theanalysis comparing hypertension and OA patients vs. normal (Table 3A)hypertension and OA patients vs. OA patients (Table 3P) and theintersection between the two populations of genes (Table 3Q).

[0070]FIG. 30 shows a venn diagram illustrating a summary of theanalysis comparing obesity and OA patients vs. normal (Table 3B) obesityand OA patients vs. OA patients (Table 3R) and the intersection betweenthe two populations of genes (Table 3S).

[0071]FIG. 31 shows a venn diagram illustrating a summary of theanalysis comparing allergy and OA patients vs. normal (Table 3C) allergyand OA patients vs. OA patients (Table 3T) and the intersection betweenthe two populations of genes (Table 3U).

[0072]FIG. 32 shows a venn diagram illustrating a summary of theanalysis comparing systemic steroids and OA patients vs. normal (Table3D) systemic steroids and OA patients vs. OA patients (Table 3V) and theintersection between the two populations of genes (Table 3W).

[0073]FIG. 33 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingManic Depression as compared with those individuals who haveSchizophrenia.

[0074]FIG. 34 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingOA and being one form of systemic steroids.

DETAILED DESCRIPTION

[0075] In accordance with the present invention, there may be employedconventional molecular biology, microbiology, and recombinant DNAtechniques within the skill of the art. Such techniques are explainedfully in the literature. See, e.g., Sambrook, Fritsch & Maniatis,“Molecular Cloning: A Laboratory Manual (1982); “DNA Cloning: APractical Approach,” Volumes I and II (D. N. Glover ed. 1985);“Oligonucleotide Synthesis” (M. J. Gait ed. 1984); “Nucleic AcidHybridization” [B. D. Hames & S. J. Higgins eds. (1985)]; “Transcriptionand Translation” [B. D. Hames & S. J. Higgins eds. (1984)]; “Animal CellCulture” [R. I. Freshney, ed. (1986)]; “Immobilized Cells And Enzymes”[IRL Press, (1986)]; B. Perbal, “A Practical Guide To Molecular Cloning”(1984). Therefore, if appearing herein, the following terms shall havethe definitions set out below.

[0076] A “cDNA” is defined as copy-DNA or complementary-DNA, and is aproduct of a reverse transcription reaction from an mRNA transcript.“RT-PCR” refers to reverse transcription polymerase chain reaction andresults in production of cDNAs that are complementary to the mRNAtemplate(s).

[0077] In addition to RT-PCR, other methods of amplifying may also beused for the purpose of measuring/quantitating tissue-specifictranscripts in human blood. For example, mass spectrometry may be usedto quantify the transcripts (Koster et al., 1996; Fu et al., 1998). Theapplication of presently disclosed method for detecting tissue-specifictranscripts in blood does not restrict to subjects undergoing course oftherapy or treatment, it may also be used for monitoring a patient forthe onset of overt symptoms of a disease. Furthermore, the presentmethod may be used for detecting any gene transcripts in blood. A kitfor diagnosing, prognosing or even predicting a disease may be designedusing gene-specific primers or probes derived from a whole blood samplefor a specific disease and applied directly to a drop of blood. A cDNAlibrary specific for a disease may be generated from whole blood samplesand used for diagnosis, prognosis or even predicting a disease.

[0078] The term “oligonucleotide” is defined as a molecule comprised oftwo or more deoxyribonucleotides and/or ribonucleotides, preferably morethan three. Its exact size will depend upon many factors which, in turn,depend upon the ultimate function and use of the oligonucleotide. Theupper limit may be 15, 20, 25, 30, 40 or 50 nucleotides in length. Theterm “primer” as used herein refers to an oligonucleotide, whetheroccurring naturally as in a purified restriction digest or producedsynthetically, which is capable of acting as a point of initiation ofsynthesis when placed under conditions in which synthesis of a primerextension product, which is complementary to a nucleic acid strand, isinduced, i.e., in the presence of nucleotides and an inducing agent suchas a DNA polymerase and at a suitable temperature and pH. The primer maybe either single-stranded or double-stranded and must be sufficientlylong to prime the synthesis of the desired extension product in thepresence of the inducing agent. The exact length of the primer willdepend upon many factors, including temperature, source of primer andthe method used. For example, for diagnostic applications, depending onthe complexity of the target sequence, the oligonucleotide primertypically contains 15-25 or more nucleotides, although it may containfewer nucleotides. The factors involved in determining the appropriatelength of primer are readily known to one of ordinary skill in the art.

[0079] As used herein, random sequence primers refer to a composition ofprimers of random sequence, i.e. not directed towards a specificsequence. These sequences possess sufficient complementary to hybridizewith a polynucleotide and the primer sequence need not reflect the exactsequence of the template.

[0080] “Restriction fragment length polymorphism” refers to variationsin DNA sequence detected by variations in the length of DNA fragmentsgenerated by restriction endonuclease digestion.

[0081] A standard Northern blot assay can be used to ascertain therelative amounts of mRNA in a cell or tissue obtained from plant orother tissue, in accordance with conventional Northern hybridizationtechniques known to those persons of ordinary skill in the art. TheNorthern blot uses a hybridization probe, e.g. radiolabelled cDNA,either containing the full-length, single stranded DNA or a fragment ofthat DNA sequence at least 20 (preferably at least 30, more preferablyat least 50, and most preferably at least 100 consecutive nucleotides inlength). The DNA hybridization probe can be labelled by any of the manydifferent methods known to those skilled in this art. The labels mostcommonly employed for these studies are radioactive elements, enzymes,chemicals which fluoresce when exposed to ultraviolet light, and others.A number of fluorescent materials are known and can be utilized aslabels. These include, for example, fluorescein, rhodamine, auramine,Texas Red, AMCA blue and Lucifer Yellow. A particular detecting materialis anti-rabbit antibody prepared in goats and conjugated withfluorescein through an isothiocyanate. Proteins can also be labelledwith a radioactive element or with an enzyme. The radioactive label canbe detected by any of the currently available counting procedures. Thepreferred isotope may be selected from ³H, ¹⁴C, ³²P, ³⁵S, ³⁶Cl, ⁵¹Cr,⁵⁷Co, ⁵⁸Co, ⁵⁹Fe, 90Y, ¹²⁵I, ¹³¹I, and ¹⁸⁶Re. Enzyme labels are likewiseuseful, and can be detected by any of the presently utilizedcolorimetric, spectrophotometric, fluorospectrophotometric, amperometricor gasometric techniques. The enzyme is conjugated to the selectedparticle by reaction with bridging molecules such as carbodiimides,diisocyanates, glutaraldehyde and the like. Many enzymes which can beused in these procedures are known and can be utilized. The preferredare peroxidase, β-glucuronidase, β-D-glucosidase, β-D-galactosidase,urease, glucose oxidase plus peroxidase and alkaline phosphatase. U.S.Pat. Nos. 3,654,090, 3,850,752, and 4,016,043 are referred to by way ofexample for their disclosure of alternate labeling material and methods.

[0082] As used herein, “individual” refers to human subjects as well asnon-human subjects. The examples herein are not meant to limit themethodology of the present invention to human subjects only, as theinstant methodology is useful in the fields of veterinary medicine,animal sciences and such. The term “individual” refers to human subjectsand non-human subjects who are disease or condition free and alsoincludes human and non-human subjects diagnosed with one or morediseases or conditions, as defined herein. “Co-morbid individuals” or“comorbidity” or “individuals considered as co-morbid” are individualswho have more than one disease or condition as defined herein. Forexample a patient diagnosed with both osteoarthritis and hypertension isconsidered to present with comorbidities.

[0083] As used herein, “detecting” refers to determining the presence ofa gene expression product, for example cDNA, RNA or EST, by any methodknown to those of skill in the art or taught in numerous texts andlaboratory manuals (see for example, Ausubel et al. Short Protocols inMolecular Biology (1995) 3rd Ed. John Wiley & Sons, Inc.). For example,methods of detection include but are not limited to, RNA fingerprinting,Northern blotting, polymerase chain reaction, ligase chain reaction,Qbeta replicase, isothermal amplification method, strand displacementamplification, transcription based amplification systems, nucleaseprotection (S1 nuclease or RNAse protection assays) as well as methodsdisclosed in WO 88/10315, WO89/06700, PCT/US87/00880, PCT/US89/01025.

[0084] As used herein, a disease of the invention includes, but is notlimited to, blood disorder, blood lipid disease, autoimmune disease,arthritis (including osteoarthritis, rheumatoid arthritis, lupus,allergies, juvenile rheumatoid arthritis and the like), bone or jointdisorder, a cardiovascular disorder (including heart failure, congenitalheart disease; rheumatic fever, valvular heart disease; corpulmonale,cardiomyopathy, myocarditis, pericardial disease; vascular diseases suchas atherosclerosis, acute myocardial infarction, ischemic heart diseaseand the like), obesity, respiratory disease (including asthma,pneumonitis, pneumonia, pulmonary infections, lung disease,bronchiectasis, tuberculosis, cystic fibrosis, interstitial lungdisease, chronic bronchitis emphysema, pulmonary hypertension, pulmonarythromboembolism, acute respiratory distress syndrome and the like),hyperlipidemias, endocrine disorder, immune disorder, infectiousdisease, muscle wasting and whole body wasting disorder, neurologicaldisorders (including migraines, seizures, epilepsy, cerebrovasculardiseases, alzheimers, dementia, Parkinson's, ataxic disorders, motorneuron diseases, cranial nerve disorders, spinal cord disorders,meningitis and the like) including neurodegenerative and/orneuropsychiatric diseases and mood disorders (including schizophrenia,anxiety, bipolar disorder; manic depression and the like, skin disorder,kidney disease, scleroderma, stroke, hereditary hemorrhagetelangiectasia, diabetes, disorders associated with diabetes (e.g.,PVD), hypertension, Gaucher's disease, cystic fibrosis, sickle cellanemia, liver disease, pancreatic disease, eye, ear, nose and/or throatdisease, diseases affecting the reproductive organs, gastrointestinaldiseases (including diseases of the colon, diseases of the spleen,appendix, gall bladder, and others) and the like. For further discussionof human diseases, see Mendelian Inheritance in Man: A Catalog of HumanGenes and Genetic Disorders by Victor A. McKusick (12th Edition (3volume set) June 1998, Johns Hopkins University Press, ISBN: 0801857422)and Harrison's Principles of Internal Medicine by Braunwald, Fauci,Kasper, Hauser, Longo, & Jameson (15th Edition, 2001), the entirety ofwhich is incorporated herein.

[0085] In another embodiment of the invention, a disease refers to animmune disorder, such as those associated with overexpression of a geneor expression of a mutant gene (e.g., autoimmune diseases, such asdiabetes mellitus, arthritis (including rheumatoid arthritis, juvenilerheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiplesclerosis, encephalomyelitis, myasthenia gravis, systemic lupuserythematosis, automimmune thyroiditis, dermatitis (including atopicdermatitis and eczematous dermatitis), psoriasis, Sjogren's Syndrome,Crohn's disease, aphthous ulcer, iritis, conjunctivitis,keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma,cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drugeruptions, leprosy reversal reactions, erythema nodosum leprosum,autoimmune uveitis, allergic encephalomyelitis, acute necrotizinghemorrhagic encephalopathy, idiopathic bilateral progressivesensorineural hearing, loss, aplastic anemia, pure red cell anemia,idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis,chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue,lichen planus, Graves' disease, sarcoidosis, primary biliary cirrhosis,uveitis posterior, and interstitial lung fibrosis), graft-versus-hostdisease, cases of transplantation, and allergy.

[0086] In another embodiment, a disease of the invention is a cellularproliferative and/or differentiative disorder that includes, but is notlimited to, cancer e.g., carcinoma, sarcoma or other metastaticdisorders and the like. As used herein, the term “cancer” refers tocells having the capacity for autonomous growth, i.e., an abnormal stateof condition characterized by rapidly proliferating cell growth.“Cancer” is meant to include all types of cancerous growths or oncogenicprocesses, metastatic tissues or malignantly transformed cells, tissues,or organs, irrespective of histopathologic type or stage ofinvasiveness. Examples of cancers include but are nor limited to solidtumors and leukemias, including: apudoma, choristoma, branchioma,malignant carcinoid syndrome, carcinoid heart disease, carcinoma (e.g.,Walker, basal cell, basosquamous, Brown-Pearce, ductal, Ehrlich tumour,in situ, Krebs 2, Merkel cell, mucinous, non-small cell lung, oat cell,papillary, scirrhous, bronchiolar, bronchogenic, squamous cell, andtransitional cell), histiocytic disorders, leukaemia (e.g., B cell,mixed cell, null cell, T cell, T-cell chronic, HTLV-II-associated,lymphocytic acute, lymphocytic chronic, mast cell, and myeloid),histiocytosis malignant, Hodgkin disease, immunoproliferative small,non-Hodgkin lymphoma, plasmacytoma, reticuloendotheliosis, melanoma,chondroblastoma, chondroma, chondrosarcoma, fibroma, fibrosarcoma, giantcell tumors, histiocytoma, lipoma, liposarcoma, mesothelioma, myxoma,myxosarcoma, osteoma, osteosarcoma, Ewing sarcoma, synovioma,adenofibroma, adenolymphoma, carcinosarcoma, chordoma,craniopharyngioma, dysgerminoma, hamartoma, mesenchymoma, mesonephroma,myosarcoma, ameloblastoma, cementoma, odontoma, teratoma, thymoma,trophoblastic tumour, adeno-carcinoma, adenoma, cholangioma,cholesteatoma, cylindroma, cystadenocarcinoma, cystadenoma, granulosacell tumour, gynandroblastoma, hepatoma, hidradenoma, islet cell tumour,Leydig cell tumour, papilloma, Sertoli cell tumour, theca cell tumour,leiomyoma, leiomyosarcoma, myoblastoma, mymoma, myosarcoma, rhabdomyoma,rhabdomyosarcoma, ependymoma, ganglioneuroma, glioma, medulloblastoma,meningioma, neurilemmoma, neuroblastoma, neuroepithelioma, neurofibroma,neuroma, paraganglioma, paraganglioma nonchromaffin, angiokeratoma,angiolymphoid hyperplasia with eosinophilia, angioma sclerosing,angiomatosis, glomangioma, hemangioendothelioma, hemangioma,hemangiopericytoma, hemangiosarcoma, lymphangioma, lymphangiomyoma,lymphangiosarcoma, pinealoma, carcinosarcoma, chondrosarcoma,cystosarcoma, phyllodes, fibrosarcoma, hemangiosarcoma, leimyosarcoma,leukosarcoma, liposarcoma, lymphangiosarcoma, myosarcoma, myxosarcoma,ovarian carcinoma, rhabdomyosarcoma, sarcoma (e.g., Ewing, experimental,Kaposi, and mast cell), neoplasms (e.g., bone, breast, digestive system,colorectal, liver, pancreatic, pituitary, testicular, orbital, head andneck, central nervous system, acoustic, pelvic respiratory tract, andurogenital), neurofibromatosis, and cervical dysplasia, and otherconditions in which cells have become immortalized or transformed.

[0087] In another embodiment, a disease of the invention includes but isnot limited to a condition wherein said condition is reflective of thestate of a particular individual, whether said state is a physical,emotional or psychological state, said state resulting from theprogression of time, treatment, environmental factors or geneticfactors.

[0088] As used herein, a gene of the invention is a gene that isexpressed in blood and is either upregulated, or downregulated and canbe used, either solely or in conjunction with other genes, as a markerfor disease as defined herein. By a gene that is expressed in blood orin a blood sample is meant a gene that is expressed in the cells whichtypically make up blood including monocytes, leukocytes, lymphocytes anderythrocytes, all other cells derived directly from haemopoietic ormesenchymal stem cells, or derived directly from a cell which typicallymakes up the blood.

[0089] The term “gene” includes a region that can be transcribed intoRNA, as the invention contemplates detection of RNA or equivalentsthereof, i.e., cDNA or EST. A gene of the invention includes but is notlimited to genes specific for or involved in a particular biologicalprocess, such as apoptosis, differentiation, stress response, aging,proliferation, etc.; cellular mechanism genes, e.g. cell-cycle, signaltransduction, metabolism of toxic compounds, and the like; diseaseassociated genes, e.g. genes involved in cancer, schizophrenia,diabetes, high blood pressure, atherosclerosis, viral-host interactionand infection and the like.

[0090] For example, the gene of the invention can be an oncogene(Hanahan, D. and R. A. Weinberg, Cell (2000) 100:57; and Yokota, J.,Carcinogenesis (2000) 21(3): 497-503) whose expression within a cellinduces that cell to become converted from a normal cell into a tumorcell. Further examples of genes of the invention include, but are notlimited to, cytokine genes (Rubinstein, M., et al., Cytokine GrowthFactor Rev. (1998) 9(2):175-81); idiotype (Id) protein genes (Benezra,R., et al., Oncogene (2001) 20(58):8334-41; Norton, J. D., J. Cell Sci.(2000) 113(22):3897-905); prion genes (Prusiner, S. B., et al., Cell(1998) 93(3):337-48; Safar, J., and S. B. Prusiner, Prog., Brain Res.,(1998) 117:421-34); genes that express molecules that induceangiogenesis (Gould, V. E. and B. M. Wagner, Hum. Pathol. (2002)33(11):1061-3); genes encoding adhesion molecules (Chothia, C. and E. Y.Jones, Annu. Rev. Biochem. (1997) 66:823-62; Parise, L. V., et al.,Semin. Cancer Biol. (2000) 10(6):407-14); genes encoding cell surfacereceptors (Deller, M. C., and Y. E. Jones, Curr. Opin. Struct. Biol.(2000) 10(2):213-9); genes of proteins that are involved inmetastasizing and/or invasive processes (Boyd, D., Cancer MetastasisRev. (1996) 15(1):77-89; Yokota, J., Carcinogenesis (2000)21(3):497-503); genes of proteases as well as of molecules that regulateapoptosis and the cell cycle (Matrisian, L. M., Curr. Biol. (1999)9(20):R776-8; Krepela, E., Neoplasma (2001) 48(5):332-49; Basbaum andWerb, Curr. Opin. Cell Biol. (1996) 8:731-738; Birkedal-Hansen, et al.,Crit. Rev. Oral Biol. Med. (1993) 4:197-250; Mignatti and Rifkin,Physiol. Rev. (1993) 73:161-195; Stetler-Stevenson, et al., Annu., Rev.Cell Biol., (1993) 9:541-573; Brinkerhoff, E., and L. M. Matrisan,Nature Reviews (2002) 3:207-214; Strasser, A., et al., Annu., Rev.Biochem., (2000), 69:217-45; Chao, D. T. and S. J. Korsmeyer, Annu. Rev.Immunol. (1998) 16:395-419; Mullauer, L., et al., Mutat. Res. (2001)488(3):211-31; Fotedar, R., et al., Prog., Cell Cycle Res., (1996),2:147-63; Reed, J. C., Am. J. Pathol., (2000) 157(5):1415-30; D'Ari, R.,Bioassays (2001) 23(7):563-5); or multi-drug resistance genes, such asMDR1 gene (Childs, S., and V. Ling, Imp., Adv. Oncol., (1994) 21-36). Inanother embodiment, a gene of the invention contains a sequence found inTables 2 or 3 or FIGS. 22-34. In another embodiment, a gene of theinvention can be an immune response gene or a non-immune response gene.By an immune response gene is meant a primary defense response genelocated outside the major histocompatibility region (MHC) that isinitially triggered in response to a foreign antigen to regulate immuneresponsiveness. All other genes expressed in blood are considered to benon-immune response gene. For example, an immune response gene would beunderstood by a person skilled in the art to include: cytokinesincluding interleukins and interferons such as TNF-alpha, IL-10, IL-12,IL-2, IL-4, IL-10, IL-12, IL-13, TGF-Beta, IFN-gamma; immunoglobulins,complement and the like (see for example Bellardelli, F. Role ofinterferons and other cytokines in the regulation of the immune responseAPMIS., 1995, March; 103(3): 161-79).

[0091] Construction of a Microarray

[0092] A nucleic acid microarray (RNA, DNA, cDNA, PCR products or ESTs)according to the invention was constructed as follows:

[0093] Nucleic acids (RNA, DNA, cDNA, PCR products or ESTs) (˜40 μl) areprecipitated with 4 μl ({fraction (1/10)} volume) of 3M sodium acetate(pH 5.2) and 10011 (2.5 volumes) of ethanol and stored overnight at −20°C. They are then centrifuged at 3,300 rpm at 4° C. for 1 hour. Theobtained pellets were washed with 50 μl ice-cold 70% ethanol andcentrifuged again for 30 minutes. The pellets are then air-dried andresuspended well in 50% dimethylsulfoxide (DMSO) or 20 μl 3×SSCovernight. The samples are then deposited either singly or in duplicateonto Gamma Amino Propyl Silane (Corning CMT-GAPS or CMT-GAP2, CatalogNo. 40003, 40004) or polylysine-coated slides (Sigma Cat. No. PO₄₂₅)using a robotic GMS 417 or 427 arrayer (Affymetrix, CA). The boundariesof the DNA spots on the microarray are marked with a diamond scriber.The invention provides for arrays where 10-20,000 different DNAs arespotted onto a solid support to prepare an array, and also may includeduplicate or triplicate DNAs. The arrays are rehydrated by suspendingthe slides over a dish of warm particle free ddH₂O for approximately oneminute (the spots will swell slightly but not run into each other) andsnap-dried on a 70-80° C. inverted heating block for 3 seconds. DNA isthen UV crosslinked to the slide (Stratagene, Stratalinker, 65 mJ—setdisplay to “650” which is 650×100 P, or baked at 80° C. for two to fourhours. The arrays are placed in a slide rack. An empty slide chamber isprepared and filled with the following solution: 3.0 grams of succinicanhydride (Aldrich) is dissolved in 189 ml of 1-methyl-2-pyrrolidinone(rapid addition of reagent is crucial); immediately after the last flakeof succinic anhydride dissolved, 21.0 ml of 0.2 M sodium borate is mixedin and the solution is poured into the slide chamber. The slide rack isplunged rapidly and evenly in the slide chamber and vigorously shaken upand down for a few seconds, making sure the slides never leave thesolution, and then mixed on an orbital shaker for 15-20 minutes. Theslide rack is then gently plunged in 95° C. ddH₂O for 2 minutes,followed by plunging five times in 95% ethanol. The slides are then airdried by allowing excess ethanol to drip onto paper towels. The arraysare then stored in the slide box at room temperature until use.

[0094] Nucleic Acid Microarrays

[0095] Any combination of the nucleic acid sequences generated fromnucleotides complimentary to regions of DNA expressed in blood are usedfor the construction of a microarray. In one embodiment, the microarrayis chondrocyte-specific and encompasses genes which are important in theosteoarthritis disease process. A microarray according to the inventionpreferably comprises between 10, 100, 500, 1000, 5000, 10,000 and 15,000nucleic acid members, and more preferably comprises at least 5000nucleic acid members. The nucleic acid members are known or novelnucleic acid sequences described herein, or any combination thereof. Amicroarray according to the invention is used to assay for differentialgene expression profiles of genes in blood samples from healthy patientsas compared to patients with a disease.

[0096] Microarray Used According to the Invention

[0097] The Human Genome U133 (HG-U133) Set, consisting of two GeneChip®arrays, contains almost 45,000 probe sets representing more than 39,000transcripts derived from approximately 33,000 well-substantiated humangenes. This set design uses sequences selected from GenBank®, dbEST, andRefSeq.

[0098] The sequence clusters were created from the UniGene database(Build 133, Apr. 20, 2001). They were then refined by analysis andcomparison with a number of other publicly available databases includingthe Washington University EST trace repository and the University ofCalifornia, Santa Cruz Golden Path human genome database (April 2001release).

[0099] The HG-U133A Array includes representation of the RefSeq databasesequences and probe sets related to sequences previously represented onthe Human Genome U95Av2 Array. The HG-U133B Array contains primarilyprobe sets representing EST clusters.

[0100] 15K ChondroChip™—The ChondroChip™ is chondrocyte-specificmicroarray chip comprising 15,000 novel and known EST sequences of thechondrocyte from human chondrocyte-specific cDNA libraries.

[0101] Controls on the ChondroChipm—There are two types of controls usedon microarrays. First, positive controls are genes whose expressionlevel is invariant between different stages of investigation and areused to monitor:

[0102] a) target DNA binding to the slide,

[0103] b) quality of the spotting and binding processes of the targetDNA onto the slide,

[0104] c) quality of the RNA samples, and

[0105] d) efficiency of the reverse transcription and fluorescentlabelling of the probes.

[0106] Second, negative controls are external controls derived from anorganism unrelated to and therefore unlikely to cross-hybridize with thesample of interest. These are used to monitor for:

[0107] a) variation in background fluorescence on the slide, and

[0108] b) non-specific hybridization.

[0109] There are currently 63 control spots on the ChondroChip™consisting of: Type No. Positive Controls: 2 Alien DNA 12 A. thalianaDNA 10 Spotting Buffer 41

[0110] BloodChip™—The “BloodChip™” is a cDNA microarray slide with10,368 PCR products derived from peripheral blood cell cDNA libraries asshown in FIG. 24.

[0111] Target Nucleic Acid Preparation and Hybridization

[0112] Preparation of Fluorescent DNA Probe from mRNA

[0113] Fluorescently labelled target nucleic acid samples are preparedfor analysis with an array of the invention. 2 μg Oligo-dT primers areannealed to 2 μg of mRNA isolated from a blood sample of a patient in atotal volume of 15 μg, by heating to 70° C. for 10 min, and cooled onice. The mRNA is reverse transcribed by incubating the sample at 42° C.for 1.5-2 hours in a 100 μg volume containing a final concentration of50 mM Tris-HCl (pH 8.3), 75 mM KCl, 3 mM MgCl₂, 25 mM DTT, 25 mMunlabelled dNTPs, 400 units of Superscript II (200 U/EL, Gibco BRL), and15 mM of Cy3 or Cy5 (Amersham). RNA is then degraded by addition of 15μl of 0.1N NaOH, and incubation at 70° C. for 10 min. The reactionmixture is neutralized by addition of 15 μl of 0.1N HCl, and the volumeis brought to 500 μl with TE (10 mM Tris, 1 mM EDTA), and 20 μg of Cotlhuman DNA (Gibco-BRL) is added.

[0114] The labelled target nucleic acid sample is purified bycentrifugation in a Centricon-30 micro-concentrator (Amicon). If twodifferent target nucleic acid samples (e.g., two samples derived from ahealthy patient vs. patient with a disease) are being analyzed andcompared by hybridization to the same array, each target nucleic acidsample is labelled with a different fluorescent label (e.g., Cy3 andCy5) and separately concentrated. The separately concentrated targetnucleic acid samples (Cy3 and Cy5 labelled) are combined into a freshcentricon, washed with 500 μl TE, and concentrated again to a volume ofless than 7 μl. 1 μl of 10 μg/μl polyA RNA (Sigma, #P9403) and 1 μl of10 μg/μl tRNA (Gibco-BRL, #15401-011) is added and the volume isadjusted to 9.5 μl with distilled water. For final target nucleic acidpreparation 2.1 μl 20×SSC (1.5M NaCl, 150 mM NaCltrate (pH8.0)) and 0.35μl 10% SDS is added.

[0115] Hybridization

[0116] Labelled nucleic acid is denatured by heating for 2 min at 100°C., and incubated at 37° C. for 20-30 min before being placed on anucleic acid array under a 22 mm×22 mm glass cover slip. Hybridizationis carried out at 65° C. for 14 to 18 hours in a custom slide chamberwith humidity maintained by a small reservoir of 3×SSC. The array iswashed by submersion and agitation for 2-5 min in 2×SSC with 0.1% SDS,followed by 1×SSC, and 0.1×SSC. Finally, the array is dried bycentrifugation for 2 min in a slide rack in a Beckman GS-6 tabletopcentrifuge in Microplus carriers at 650 RPM for 2 min.

[0117] Signal Detection and Data Generation

[0118] Following hybridization of an array with one or more labelledtarget nucleic acid samples, arrays are scanned immediately using a GMSScanner 418 and Scanalyzer software (Michael Eisen, StanfordUniversity), followed by GeneSpring™ software (Silicon Genetics, CA)analysis. Alternatively, a GMS Scanner 428 and Jaguar software may beused followed by GeneSpring™ software analysis.

[0119] If one target nucleic acid sample is analyzed, the sample islabelled with one fluorescent dye (e.g., Cy3 or Cy5).

[0120] After hybridization to a microarray as described herein,fluorescence intensities at the associated nucleic acid members on themicroarray are determined from images taken with a custom confocalmicroscope equipped with laser excitation sources and interferencefilters appropriate for the Cy3 or Cy5 fluorescence.

[0121] The presence of Cy3 or Cy5 fluorescent dye on the microarrayindicates hybridization of a target nucleic acid and a specific nucleicacid member on the microarray. The intensity of Cy3 or Cy5 fluorescencerepresents the amount of target nucleic acid which is hybridized to thenucleic acid member on the microarray, and is indicative of theexpression level of the specific nucleic acid member sequence in thetarget sample.

[0122] After hybridization, fluorescence intensities at the associatednucleic acid members on the microarray are determined from images takenwith a custom confocal microscope equipped with laser excitation sourcesand interference filters appropriate for the Cy3 and Cy5 fluors.Separate scans are taken for each fluor at a resolution of 225 m2 perpixel and 65,536 gray levels. Normalization between the images is usedto adjust for the different efficiencies in labeling and detection withthe two different fluors. This is achieved by manual matching of thedetection sensitivities to bring a set of internal control genes tonearly equal intensity followed by computational calculation of theresidual scalar required for optimal intensity matching for this set ofgenes.

[0123] The presence of Cy3 or Cy5 fluorescent dye on the microarrayindicates hybridization of a target nucleic acid and a specific nucleicacid member on the microarray. The intensities of Cy3 or Cy5fluorescence represent the amount of target nucleic acid which ishybridized to the nucleic acid member on the microarray, and isindicative of the expression level of the specific nucleic acid membersequence in the target sample. If a nucleic acid member on the arrayshows no color, it indicates that the gene in that element is notexpressed in either sample. If a nucleic acid member on the array showsa single color, it indicates that a labelled gene is expressed only inthat cell sample. The appearance of both colors indicates that the geneis expressed in both tissue samples. The ratios of Cy3 and Cy5fluorescence intensities, after normalization, are indicative ofdifferences of expression levels of the associated nucleic acid membersequence in the two samples for comparison. A ratio of expression notequal to is used as an indication of differential gene expression.

[0124] The array is scanned in the Cy 3 and Cy5 channels and stored asseparate 16-bit TIFF images. The images are incorporated and analyzedusing Scanalyzer software which includes a gridding process to capturethe hybridization intensity data from each spot on the array. Thefluorescence intensity and background-subtracted hybridization intensityof each spot is collected and a ratio of measured mean intensities ofCy5 to Cy3 is calculated. A liner regression approach is used fornormalization and assumes that a scatter plot of the measured Cy5 versusCy3 intensities should have a scope of one. The average of the ratios iscalculated and used to rescale the data and adjust the slope to one. Apost-normalization cutoff of a ratio not equal to 1.0-is used toidentify differentially expressed genes.

[0125] When comparing two or more samples for differences, results arereported as statistically significant when there is only a smallprobability that similar results would have been observed if the testedhypothesis (i.e., the genes are not expressed at different levels) weretrue. A small probability can be defined as the accepted threshold levelat which the results being compared are considered significantlydifferent. The accepted lower threshold is set at, but not limited to,0.05 (i.e., there is a 5% likelihood that the results would be observedbetween two or more identical populations) such that any valuesdetermined by statistical means at or below this threshold areconsidered significant.

[0126] When comparing two or more samples for similarities, results arereported as statistically significant when there is only a smallprobability that similar results would have been observed if the testedhypothesis (i.e., the genes are not expressed at different levels) weretrue. A small probability can be defined as the accepted threshold levelat which the results being compared are considered significantlydifferent. The accepted lower threshold is set at, but not limited to,0.05 (i.e., there is a 5% likelihood that the results would be observedbetween two or more identical populations) such that any valuesdetermined by statistical means above this threshold are not consideredsignificantly different and thus similar.

[0127] Identification of genes differentially expressed in blood samplesfrom patients with disease as compared to healthy patients or ascompared to patients without said disease is determined by statisticalanalysis of the gene expression profiles from healthy patients orpatients without disease compared to patients with disease using theWilcox Mann Whitney rank sum test. Other statistical tests can also beused, see for example (Sokal and Rohlf (1987) Introduction toBiostatistics 2nd edition, WH Freeman, New York), which is incorporatedherein in their entirety.

[0128] In order to facilitate ready access, e.g. for comparison, review,recovery and/or modification, the expression profiles of patients withdisease and/or patients without disease or healthy patients can berecorded in a database, whether in a relational database accessible by acomputational device or other format, or a manually accessible indexedfile of profiles as photographs, analogue or digital imaging, readoutsspreadsheets etc. Typically the database is compiled and maintained at acentral facility, with access being available locally and/or remotely.

[0129] As would be understood by a person skilled in the art, comparisonas between the expression profile of a test patient with expressionprofiles of patients with a disease, expression profiles of patientswith a certain stage or degree of progression of said disease, withoutsaid disease, or a healthy patient so as to diagnose or prognose saidtest patient can occur via expression profiles generated concurrently ornon concurrently. It would be understood that expression profiles can bestored in a database to allow said comparison.

[0130] As additional test samples from test patients are obtained,through clinical trials, further investigation, or the like, additionaldata can be determined in accordance with the methods disclosed hereinand can likewise be added to a database to provide better reference datafor comparison of healthy and/or non-disease patients and/or certainstage or degree of progression of a disease as compared with the testpatient sample.

[0131] Use of Expression Profiles for Diagnostic Purposes

[0132] As would be understood to a person skilled in the art, one canutilize sets of genes which have been identified as statisticallysignificant as described above in order to characterize an unknownsample as having said disease or not having said disease. This iscommonly termed “class prediction”.

[0133] Methods that can be used for class prediction analysis have beenwell described and generally involve a training phase using samples withknown classification and a testing phase from which the algorithmgeneralizes from the training data so as to predict classification ofunknown samples (see for Example Slonim, D. (2002), Nature GeneticsSupp., Vol. 32 502-8, Raychaudhuri et al., (2001) Trends Biotechnol.,19: 189-193; Khan et al. (2001) Nature Med., 7 673-9; Golub et al.(1999) Science 286: 531-7. Hastie et al., (2000) Genome Biol., 1(2)Research 0003.1-0003.21, all of which are incorporated herein byreference in their entirety).

[0134] As additional samples are obtained, for example during clinicaltrials, their expression profiles can be determined and correlated withthe relevant subject data in the database and likewise be recorded insaid database. Algorithms as described above can be used to queryadditional samples against the existing database to further refine thediagnostic and/or prognostic determination by allowing an even greaterassociation between the disease and gene expression signature.

[0135] The diagnosing or prognosing may thus be performed by detectingthe expression level of two or more genes, three or more genes, four ormore genes, five or more genes, six or more genes, seven or more genes,eight or more genes, nine or more genes, ten or more genes, fifteen ormore genes, twenty or more genes thirty or more genes, fifty or moregenes, one hundred or more genes, two hundred or more genes, threehundred or more genes, five hundred or more genes or all of the genesdisclosed for the specific disease in question.

[0136] Data Acquisition and Analysis of differentially Expressed ESTSequences

[0137] The differentially expressed EST sequences are then searchedagainst available databases, including the “nt”, “nr”, “est”, “gss” and“htg” databases available through NCBI to determine putative identitiesfor ESTs matching to known genes or other ESTs. Functionalcharacterisation of ESTs with known gene matches are made according toany known method. Preferably, differentially expressed EST sequences arecompared to the non-redundant Genbank/EMBL/DDBJ and dbEST databasesusing the BLAST algorithm (Altschul S F, Gish W, Miller W, Myers E W,Lipman D J., Basic local alignment search tool., J. Mol. Biol., 1990;215:403-10). A minimum value of P=10¹⁰ and nucleotide sequenceidentity>95%, where the sequence identity is non-contiguous orscattered, are required for assignments of putative identities for ESTsmatching to known genes or to other ESTs. Construction of anon-redundant list of genes represented in the EST set is done with thehelp of Unigene, Entrez and PubMed at the National Center forBiotechnology Information (NCBI) web site at www.ncbi.nlm.nih.gov.

[0138] Genes are identified from ESTs according to known methods. Toidentify novel genes from an EST sequence, the EST should preferably beat least 100 nucleotides in length, and more preferably 150 nucleotidesin length, for annotation. Preferably, the EST exhibits open readingframe characteristics (i.e., can encode a putative polypeptide).

[0139] Because of the completion of the Human Genome Project, a specificEST which matches with a genomic sequence can be mapped onto a specificchromosome based on the chromosomal location of the genomic sequence.However, no function may be known for the protein encoded by thesequence and the EST would then be considered “novel” in a functionalsense. In one aspect, the invention is used to identify a noveldifferentially expressed EST, which is part of a larger known sequencefor which no function is known, is used to determine the function of agene comprising the EST. Alternatively, or additionally, the EST can beused to identify an mRNA or polypeptide encoded by the larger sequenceas a diagnostic or prognostic marker of a disease.

[0140] Having identified an EST corresponding to a larger sequence,other portions of the larger sequence which comprises the EST can beused in assays to elucidate gene function, e.g., to isolate polypeptidesencoded by the gene, to generate antibodies specifically reactive withthese polypeptides, to identify binding partners of the polypeptides(receptors, ligands, agonists, antagonists and the like) and/or todetect the expression of the gene (or lack thereof) in healthy ordiseased individuals.

[0141] In another aspect, the invention provides for nucleic acidsequences that do not demonstrate a “significant match” to any of thepublicly known sequences in sequence databases at the time a query isdone. Longer genomic segments comprising these types of novel ESTsequences can be identified by probing genomic libraries, while longerexpressed sequences can be identified in cDNA libraries and/or byperforming polymerase extension reactions (e.g., RACE) using ESTsequences to derive primer sequences as is known in the art. Longerfragments can be mapped to particular chromosomes by FISH and othertechniques and their sequences compared to known sequences in genomicand/or expressed sequence databases.

[0142] The amino acid sequences encoded by the ESTs can also be used tosearch databases, such as GenBank, SWISS-PROT, EMBL database, PIRprotein database, Vecbase, or GenPept for the amino acid sequences ofthe corresponding full-length genes according to procedures well knownin the art.

[0143] Identified genes can be catalogued according to their putativefunction. Functional characterization of ESTs with known gene matches ispreferably made according to the categories described by Hwang et alCompendium of Cardiovascular Genes. Circulation 1997;96:4146-203. Thedistribution of genes in each of the subcellular categories will provideimportant insights into the disease process.

[0144] Alternative methods for analysing ESTs are also available. Forexample, the ESTs may be assembled into contigs with sequence alignment,editing, and assembly programs such as PHRED and PHRAP (Ewing, et al.,1998, Genome Res., 3:175, incorporated herein; and the web site atbozeman.genome.washington.edu). Contig redundancy is reduced byclustering nonoverlapping sequence contigs using the EST cloneidentification number, which is common for the nonoverlapping 5 and 3sequence reads for a single EST cDNA clone. In one aspect, the consensussequence from each cluster is compared to the non-redundantGenbank/EMBL/DDBJ and dbEST databases using the BLAST algorithm with thehelp of unigene, Entrez and PubMed at the NCBI site.

[0145] Known Nucleic Acid Sequences or ESTs and Novel Nucleic AcidSequences or ESTs

[0146] An EST that exhibits a significant match (>65%, and preferably90% or greater, identity) to at least one existing sequence in anexisting nucleic acid sequence database is characterised as a “known”sequence according to the invention. Within this category, some knownESTs match to existing sequences which encode polypeptides with knownfunction(s) and are referred to as a “known sequence with a function”.Other “known” ESTs exhibit a significant match to existing sequenceswhich encode polypeptides of unknown function(s) and are referred to asa “known sequence with no known function”.

[0147] EST sequences which have no significant match (less than 65%identity) to any existing sequence in the above cited availabledatabases are categorised as novel ESTs. To identify a novel gene froman EST sequence, the EST is preferably at least 150 nucleotides inlength. More preferably, the EST encodes at least part of an openreading frame, that is, a nucleic acid sequence between a translationinitiation codon and a termination codon, which is potentiallytranslated into a polypeptide sequence.

[0148] The following references were cited herein:

[0149] Claudio J O et al. (1998). Genomics 50:44-52.

[0150] Chelly J et al. (1989). Proc. Nat. Acad. Sci. USA. 86:2617-2621.

[0151] Chelly J et al. (1988). Nature 333:858-860.

[0152] Drews J & Ryser S (1997). Nature Biotech. 15:1318-9.

[0153] Ferrie R M et al. (1992). Am. J. Hum. Genet. 51:251-62.

[0154] Fu D-J et al. (1998). Nat. Biotech 16: 381-4.

[0155] Gala J L et al. (1998). Clin. Chem. 44(3):472-81.

[0156] Geisterfer-Lowrance A AT et al. (1990). Cell 62:999-1006.

[0157] Groden J et al. (1991). Cell 66:589-600.

[0158] Hwang D M et al. (1997). Circulation 96:4146-4203.

[0159] Jandreski M A & Liew C C (1987). Hum. Genet. 76:47-53.

[0160] Jin 0 et al. (1990). Circulation 82:8-16

[0161] Kimoto Y (1998). Mol. Gen. Genet 258:233-239.

[0162] Koster M et al. (1996). Nat. Biotech 14: 1123-8.

[0163] Liew & Jandreski (1986). Proc. Nat. Acad. Sci. USA. 83:3175-3179

[0164] Liew C C et al. (1990). Nucleic Acids Res. 18:3647-3651.

[0165] Liew C C (1993). J. Mol. Cell. Cardiol. 25:891-894

[0166] Liew C C et al. (1994). Proc. Natl. Acad. Sci. USA.91:10645-10649.

[0167] Liew et al. (1997). Mol. and Cell. Biochem. 172:81-87.

[0168] Niimura H et al. (1998). New Eng. J. Med. 338:1248-1257.

[0169] Ogawa M (1993). Blood 81:2844-2853.

[0170] Santoro I M & Groden J (1997). Cancer Res. 57:488-494.

[0171] Yuasa T et al. (1998). Japanese J Cancer Res. 89:879-882.

[0172] Description of Tables:

[0173] Table 1: Overlap of Genes Expressed in Blood

[0174] (Estimated from about 5,100 unique known genes from the over25,000 ESTs obtained from human blood cDNA libraries).

[0175] Table 2: Comparison of approximately 5,140 Unique GenesIdentified in the Blood Cell cDNA Library to Genes Previously Identifiedin Specific Tissues

[0176] Column 1: List of unique genes derived from 25,000 known ESTsfrom blood cells.

[0177] Column 2: Number of genes found in randomly sequenced ESTs fromblood cells.

[0178] Column 3: Accession number.

[0179] Column 4: “+” indicates the presence of the unique gene inpublicly available cDNA libraries of blood (Bl), brain (Br), heart (H),kidney (K), liver (Li) and lung (Lu). **Comparison to previouslyidentified tissue-specific genes was determined using the GenBank of theNational Centre of Biotechnology Information (NCBI) Database.

[0180] Table 3 shows genes that are differentially expressed in bloodsamples from patients with different diseases as compared to bloodsamples from healthy patients.

[0181] Table 3A shows the identity of those genes that aredifferentially expressed in blood samples from patients withosteoarthritis and hypertension as compared with normal patients asdepicted in FIG. 8

[0182] Table 3B shows the identity of those genes that aredifferentially expressed in blood samples from patients withosteoarthritis and obesity as compared with normal patients as depictedin FIG. 9.

[0183] Table 3C shows the identity of those genes that aredifferentially expressed in blood samples from patients withosteoarthritis and allergies as compared with normal patients asdepicted in FIG. 10.

[0184] Table 3D shows the identity of those genes that aredifferentially expressed in blood samples from patients withosteoarthritis and subject to systemic steroids as compared with normalpatients as depicted in FIG. 11.

[0185] Table 3E shows the identity of those genes that aredifferentially expressed in blood samples from patients withhypertension as depicted in FIG. 12.

[0186] Table 3F shows the identity of those genes that aredifferentially expressed in blood samples from patients obesity asdepicted in FIG. 13.

[0187] Table 3G shows the identity of those genes that aredifferentially expressed in blood samples from patients with type IIdiabetes as depicted in FIG. 14.

[0188] Table 3H shows the identity of those genes that aredifferentially expressed in blood samples from patients withhyperlipidemia as depicted in FIG. 15.

[0189] Table 31 shows the identity of those genes that aredifferentially expressed in blood samples from patients with lungdisease as depicted in FIG. 16.

[0190] Table 3J shows the identity of those genes that aredifferentially expressed in blood samples from patients with bladdercancer as depicted in FIG. 17.

[0191] Table 3K shows the identity of those genes that aredifferentially expressed in blood samples from patients with bladdercancer as depicted in FIG. 18.

[0192] Table 3L shows the identity of those genes that aredifferentially expressed in blood samples from patients with coronaryartery disease (CAD) as depicted in FIG. 19.

[0193] Table 3M shows the identity of those genes that aredifferentially expressed in blood samples from patients with rheumatoidarthritis as depicted in FIG. 20.

[0194] Table 3N shows the identity of those genes that aredifferentially expressed in blood samples from patients with depressionas depicted in FIG. 21.

[0195] Table 30 shows the identity of those genes that aredifferentially expressed in blood samples from patients with variousstages of osteoarthritis as depicted in FIG. 22.

[0196] Table 3P shows the identity of those genes that aredifferentially expressed in blood samples from patients withhypertension and OA when compared with patients who have OA only whereingenes identified in Table 3A have been removed so as to identify geneswhich are unique to hypertension.

[0197] Table 3Q shows the identity of those genes which were identifiedin Table 3A which are shared with those genes differentially expressedin blood samples from patients with hypertension and OA when comparedwith patients who have OA only.

[0198] Table 3R shows the identity of those genes that aredifferentially expressed in blood samples from patients who are obeseand have OA when compared with patients who have OA only and whereingenes identified in Table 3B have been removed so as to identify geneswhich are unique to obesity.

[0199] Table 3S shows the identify of those genes identified in Table 3Bwhich are shared with those genes differentially expressed in bloodsamples from patients who are obese and have OA when compared withpatients who have OA.

[0200] Table 3T shows the identity of those genes that aredifferentially expressed in blood samples from patients with allergiesand OA when compared with patients who have OA only wherein genesidentified in Table 3C have been removed so as to identify genes whichare unique to allergies.

[0201] Table 3U shows the identify of those genes identified in Table 3Cwhich are shared with those genes differentially expressed in bloodsamples from patients with allergies and OA when compared with patientswho have OA only.

[0202] Table 3V shows the identity of those genes that aredifferentially expressed in blood samples from patients who are onsystemic steroids and have OA when compared with patients who have OAonly wherein genes identified in Table 3D have been removed so as toidentify genes which are unique to patients on systemic steroids.

[0203] Table 3W shows the identify of those genes identified in Table 3Dwhich are shared with those genes differentially expressed in bloodsamples from patients who are on systemic steroids and have OA whencompared with patients who have OA only.

[0204] Table 3X shows the identity of those genes that aredifferentially expressed in blood samples from patients with livercancer as depicted in FIG. 25.

[0205] Table 3Y shows the identity of those genes that aredifferentially expressed in blood samples from patients withschizophrenia as depicted in FIG. 26.

[0206] Table 3Z shows the identity of those genes that aredifferentially expressed in blood samples from patients with Chagasdisease as depicted in FIG. 27.

[0207] Table 3AA shows the identity of those genes that aredifferentially expressed in blood samples from patients with asthma asdepicted in FIG. 28.

[0208] Table 3AB shows the identity of those genes that aredifferentially expressed in blood from patients with either mild orsevere OA, but for which genes relevant to asthma, obesity,hypertension, systemic steroids and allergies have been removed.

[0209] Table 3AC shows the identity of those genes that aredifferentially expressed in blood from patients with schizophrenia ascompared with manic depression syndrome (MDS).

[0210] Table 3AD shows the identity of those genes that aredifferentially expressed in blood from patients taking either birthcontrol, prednisone or hormone replacement therapy and presenting withOA as depicted in FIG. 34.

[0211] Table 4 shows 102 EST sequences of Tables 3A-3AD with“no-significant match” to known gene sequences.

[0212] Table 5 shows a list of genes showing greater than two folddifferential expression in CAD peripheral blood cells vs. normal bloodcells.

[0213] The following examples are given for the purpose of illustratingvarious embodiments of the invention and are not meant to limit thepresent invention in any fashion.

EXAMPLE 1

[0214] Construction of a cDNA Library

[0215] RNA extracted from human tissues (including fetal heart, adultheart, liver, brain, prostate gland and whole blood) were used toconstruct unidirectional cDNA libraries. The first mammalian heart cDNAlibrary was constructed as early as 1982. Since then, the methodologyhas been revised and optimal conditions have been developed forconstruction of human heart and hematopoietic progenitor cDNA libraries(Liew et al., 1984; Liew 1993, Claudio et al., 1998). Most of the novelgenes which were identified by sequence annotation can now be obtainedas full length transcripts.

EXAMPLE 2

[0216] Catalogue of EST Database

[0217] Random partial sequencing of expressed sequence tags (ESTs) ofcDNA clones from the blood cell library was carried out to establish anEST database of blood. The known genes as derived from the ESTs werecategorized into seven major cellular functions (Hwang, Dempsey et al.,1997). The preparation of the chondrocyte-specific EST database isreported in WO 02/070737, which is hereby incorporated by reference inits entirety.

EXAMPLE 3

[0218] Differential Screening of cDNA Library

[0219] cDNA probes generated from transcripts of each tissue were usedto hybridize the blood cell cDNA clones or chondrocyte cDNA clones (Liewet al., 1997; WO 02/070737). The “positive” signals which werehybridized with P-labelled cDNA probes were defined as genes whichshared identity with blood and respective tissues. The “negative” spotswhich were not exposed to P-labelled cDNA probes were considered to beblood-cell-enriched or low frequency transcripts.

EXAMPLE 4

[0220] Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) Assay

[0221] RNA extracted from samples of human tissue was used for RT-PCRanalysis (Jin et al. 1990). Three pairs of forward and reverse primerswere designed for human cardiac beta-myosin heavy chain gene (βMyHC),amyloid precursor protein (APP) gene and adenomatous polyposis-coliprotein (APC) gene. The PCR products were also subjected to automatedDNA sequencing to verify the sequences as derived from the specifictranscripts of blood.

EXAMPLE 5

[0222] Detection of Tissue Specific Gene Expression in Human Blood UsingRT-PCR

[0223] The beta-myosin heavy chain gene (βMyHC) transcript (mRNA) isknown to be highly expressed in ventricles of the human heart. Thissarcomeric protein is important for heart muscle contraction and itspresence would not be expected in other non-muscle tissues and blood. In1990, the gene for human cardiac βMyHC was completely sequenced (Liew etal. 1990) and was comprised of 41 exons and 42 introns.

[0224] The method of reverse transcription polymerase chain reaction(RT-PCR) was used to determine whether this cardiac specific mRNA isalso present in human blood. A pair of primers was designed; the forwardprimer (SEQ ID No. 3) was on the boundary of exons 21 and 22, and thereverse primer (SEQ ID No. 4) was on the boundary of exons 24 and 25.This region of mRNA is only present in PMyHC and is not found in thealpha-myosin heavy chain gene (aMyHC).

[0225] A blood sample was first treated with lysing buffer and thenundergone centrifuge. The resulting pellets were further processed withRT-PCR. RT-PCR was performed using the total blood cell RNA as atemplate. A nested PCR product was generated and used for sequencing.The sequencing results were subjected to BLAST and the identity of exons21 to 25 was confirmed to be from βMyHC (FIG. 1A).

[0226] Using the same method just described, two other tissue specificgenes—amyloid precursor protein (APP, forward primer, SEQ ID No. 7;reverse primer, SEQ ID No. 8) found in the brain and associated withAlzheimer's disease, and adenomatous polyposis coli protein (APC) foundin the colon and rectum and associated with colorectal cancer (Groden etal. 1991; Santoro and Groden 1997)-were also detected in the RNAextracted from human blood (FIG. 1B).

EXAMPLE 6

[0227] Multiple RT-PCR Analysis on a Drop of Blood from aNormal/Diseased Individual

[0228] A drop of blood was extracted to obtain RNA to carry outquantitative RT-PCR analysis. Specific primers for the insulin gene weredesigned: forward primer (5′-GCCCTCTGGGGACCTGAC-3′, SEQ ID NO 1) of exon1 and reverse primer (5′-CCCACCTGCAGGTCCTCT-3″, SEQ ID NO 2) of exons 1and 2 of insulin gene. Such reverse primer was obtained by deleting theintron between the exons 1 and 2. Blood samples of 4 normal subjectswere assayed. It was found that the insulin gene is expressed in theblood and the quantitative expression of the insulin gene in a drop ofblood is influenced by fasting and non-fasting states of normal healthysubjects (FIG. 2). This very low level of expression of the insulin genereflects the phenotypic status of a person and strongly suggests thatthere is a physiological and pathological role for its expression,contrary to the basal or illegitimate theory of transcription suggestedby Chelly et al. (1989) and Kimoto (1998).

[0229] Same quantitative RT-PCR analysis was performed using insulinspecific primers on RNA samples extracted from a drop of blood from anormal healthy person, a person having late-onset diabetes (Type II) anda person having asymptomatic diabetes. It was found that the insulingene is expressed differentially amongst subjects that are healthy,diagnosed as type II diabetic, and also in an asymptomatic preclinicalpatient (FIG. 3).

[0230] Similarly, specific primers for the atrial natriuretic factor(ANF) gene were designed (forward primer, SEQ ID No. 5; reverse primer,SEQ ID No. 6) and RT-PCR analysis was performed on a drop of blood. ANFis known to be highly expressed in heart tissue biopsies and in theplasma of heart failure patients. However, atrial natriuretic factor wasobserved to be expressed in the blood and the expression of the atrialnatriuretic factor gene is significantly higher in the blood of patientswith heart failure as compared to the blood of a normal control patient.

[0231] Specific primers for the zinc finger protein gene (ZFP, forwardprimer, SEQ ID No. 9; reverse primer, SEQ ID No. 10) were also designedand RT-PCR analysis was performed on a drop of blood. ZFP is known to behigh in heart tissue biopsies of cardiac hypertrophy and heart failurepatients. In the present study, the expression of ZFP was observed inthe blood as well as differential expression levels of ZFP amongst thenormal, diabetic and asymptomatic preclinical subjects (FIG. 4);although neither of the non-normal subjects has been specificallydiagnosed as suffering from cardiac hypertrophy and/or heart failure,the higher expression levels of the ZFP gene in their blood may indicatethat these subjects are headed in that general direction.

[0232] It was hypothesized that a housekeeping gene such asglyceraldehyde dehydrogenase (GADH) which is required and highlyexpressed in all cells would not be differentially expressed in theblood of normal vs. disease subjects. This hypothesis was confirmed byRT-PCR using GADH specific primers (FIG. 4). Thus, GADH is useful as aninternal control.

[0233] Standardized levels of insulin gene or ZFP gene expressed in adrop of blood were estimated using a housekeeping gene as an internalcontrol relative to insulin or ZFP expressed (FIGS. 5A & 5B). The levelsof insulin gene expressed in each fractionated cell from whole bloodwere also standardized and shown in FIG. 5C.

EXAMPLE 7

[0234] Human Blood Cell cDNA Library

[0235] In order to further substantiate the present invention,differential screening of the human blood cell cDNA library wasconducted. cDNA probes derived from human blood, adult heart or brainwere respectively hybridized to the human blood cDNA library clones. Asshown in FIG. 7, more than 95% of the “positively” identified clones areidentical between the blood and other tissue samples.

[0236] DNA sequencing of randomly selected clones from the human wholeblood cell cDNA library was also performed. This allowed informationregarding the cellular function of blood to be obtained concurrentlywith gene identification. More than 20,000 expressed sequence tags(ESTs) have been generated and characterized to date, 17.6% of which didnot result in a statistically significant match to entries in theGenBank databases and thus were designated as “Novel” ESTs. Theseresults are summarized in FIG. 7 together with the seven cellularfunctions related to percent distribution of known genes in blood and inthe fetal heart.

[0237] From 20,000 ESTs, 1,800 have been identified as known genes whichmay not all appear in the hemapoietic system. For example, the insulingene and the atrial natriuretic factor gene have not been detected inthese 20,000 ESTs but their transcripts were detected in a drop ofblood, strongly suggesting that all transcripts of the human genome canbe detected by performing RT-PCR analysis on a drop of blood.

[0238] In addition, approximately 400 novel genes have been identifiedfrom the 20,000 ESTs characterized to date, and these will be subjectedto full length sequencing and open reading frame alignment to reduce theactual number of novel ESTs prior to screening for disease markers.

[0239] Analysis of the approximately 6,283 ESTs which have known matchesin the GenBank databases revealed that this dataset represents over1,800 unique genes. These genes have been catalogued into seven cellularfunctions. Comparisons of this set of unique genes with ESTs derivedfrom human brain, heart, lung and kidney demonstrated a greater than 50%overlap in expression (Table 1). TABLE 1 Overlap of Genes Expressed inBlood Tissue UniGene* Overlap Brain 19,158 70% Heart 17,021 67% Kidney19,414 69% Liver 22,836 71% Lung 22,209 75%

[0240] There are about 5,100 unique known genes from the over 25,000ESTs obtained from human blood cDNA libraries. These genes were searchedagainst human UniGene, Build #160 (with a total of 111,064 clusters).

EXAMPLE 8

[0241] Blood cell ESTs

[0242] The results from the differential screening clearly indicate thatthe transcripts expressed in the whole blood are reflective of genesexpressed in all cells and tissues of the body. More than 95% ofdetectable spots were identical from two different tissues. Theremaining 5% of spots may represent cell- or tissue-specifictranscripts; however, results obtained from partial sequencing togenerate ESTs of these clones revealed most of them not to be cell- ortissue-specific transcripts. Therefore, the negative spots arepostulated to be reflective of low abundance transcripts in the tissuefrom which the cDNA probes were derived.

[0243] An alternative approach that was employed to identify transcriptsexpressed at low levels is the large-scale generation of expressedsequence tags (ESTs). There is substantial evidence regarding theefficiency of this technology to detect previously characterized (known)and uncharacterized (unknown or novel) genes expressed in thecardiovascular system (Hwang & Dempsey et al. 1997). In the presentinvention, 20,000 ESTs have been produced from a human blood cell cDNAlibrary and resulted in the identification of approximately 1,800 uniqueknown genes (Table 2)

[0244] In the most recent GenBank release, analysis of more than 300,000ESTs in the database (dbESTs) generated more than 48,000 gene clusterswhich are thought to represent approximately 50% of the genes in thehuman genome. Only 4,800 of the dbESTs are blood-derived. In the presentinvention, 20,000 ESTs have been obtained to date from a human bloodcDNA library, which provides the world's most informative database withrespect to blood cell transcripts. From the limited amount ofinformation generated so far (i.e. 1,800 unique genes), it has alreadybeen determined that more than 50% of the transcripts are found in othercells or tissues of the human body (Table 2). Thus, it is expected thatby increasing the number of ESTs generated, more genes will beidentified that have an overlap in expression between the blood andother tissues. Furthermore, the transcripts for several genes which areknown to have tissue-restricted patterns of expression (i.e. βMyHC, APP,APC, ANF, ZFP) have also been demonstrated to be present in blood.

[0245] Most recently, a cDNA library of human hematopoietic progenitorstem cells has also been constructed. From the limited set of 1,000ESTs, there are at least 200 known genes that are shared with othertissue related genes (Claudio et al. 1998).

[0246] Table 2 demonstrates the expression of known genes of specifictissues in blood cells. Previously, only the presence of “housekeeping”genes would have been expected. Additionally, the presence of at least25 of the currently known 500 genes corresponding to molecular drugtargets was detected. These molecular drug targets are used in thetreatment of a variety of diseases which involve inflammation, renal andcardiovascular function, neoplastic disease, immunomodulation and viralinfection (Drews & Ryser, 1997). It is expected that additional novelESTs will represent future molecular drug targets.

EXAMPLE 9

[0247] Blood cDNA chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having coronary arterydisease as compared with gene expression profiles from normalindividuals.

[0248] A microarray was constructed using cDNA clones from a humanperipheral blood cell cDNA library, as described herein. A total of10,368 polymerase chain reaction (PCR) products of the clones from thehuman peripheral blood cell cDNA library described herein were arrayedusing GNS 417 arrayer (Affymetrix). RNA for microarray analysis wasisolated from whole blood samples obtained from three male and onefemale patients with coronary heart disease (80-90% stenosis) receivingvascular extension drugs and awaiting bypass surgery, and three healthymale controls.

[0249] A method of high-fidelity mRNA amplification from 1 pg of totalRNA sample was used. Cy5- or Cy3-dUTP was incorporated into cDNA probesby reverse transcription of anti-sense RNA, primed by oligo-dT. Labelledprobes were purified and concentrated to the desired volume.Pre-hybridization and hybridization were performed following Hegde'sprotocol (Hegde P et al., A concise guide to cDNA microarray analysis.Biotechniques, 2000; 29: 548-56). After overnight hybridization andwashing, hybridization signals were detected with a GMS 418 scanner at635-nm (Cy5) and 532-nm (Cy3) wave lengths (see FIG. 24). Two RNA poolswere labelled alternatively with Cy5- and Cy3-dUTP, and each experimentwas repeated twice. Cluster analysis using GeneSpring™ 4.1.5 (SiliconGenetics) revealed two distinct groups consisting of four CAD and threenormal control samples. Two images scanned at different wavelengths weresuper-imposed. Individual spots were identified on a customized grid. Of10,368 spots, 10,012 (96.6%) were selected after the removal of spotswith irregular shapes. Data quality was assessed with values of ChlGTB2and Ch2GTB2 provided by ScanAlyze. Only spots with ChlGTB2 and Ch2GTB2over 0.50 were selected. After evaluation of signal intensities, 8750(84.4%) spots were left. Signal intensities were normalized using ascatter-plot of the signal intensities of the two channels. Afternormalization, the expression ratios of α-actin were 1.00+0 21,1.11+0.22, 1.14+0.20 and 1.30+0.18 (24 samples of α-actin were spottedon this slide as the positive control) in the four images. Genedifferential expression was assessed as the ratio of two wave-lengthsignal intensities. Spots showing a differential expression more thantwofold in all four experiments were identified as peripheral bloodcell, differentially expressed candidate genes in CAD. 108 genes aredifferentially expressed in CAD peripheral blood cells. 43 genes aredown-regulated in CAD blood cells and 65 are upregulated (see Table 5).Functional characterization of these genes shows that differentialexpression takes place in every gene functional category, indicatingthat profound changes occur in CAD blood cells.

[0250] The differential expression of three genes, pro-platelet basicprotein (PBP), platelet factor 4 (PF4) and coagulation factor XIII A1 (F13A), initially identified in the microarray data analysis, was furtherexamined by reverse transcriptase-PCR (RT-PCR) using the Titan One-tubeRT-PCR kit (Boehringer Mannheim). Reaction solution contains 0.2 mM eachdNTP, 5 mM DTT, 1.5 mM MgCl 0.1 pg of total RNA from each sample and 20pmol each of left and right primers of PBP (5′-GGTGCTGCTGCTTCTGTCAT-3′and 5′-GGCAGATTTT CCTCCCATCC-3′), F13A (5′-AGTCCACCGTGCTAACCATC-3′ and5′-AGGGAGTCACTGCTCATGCT-3′) and PF4 (5′ GTTGCTGCTCCTGCCACTT 3′ and 5′GTGGCTATCAGTTGGGCAGT-3′). RT-PCR steps are as follows: 1.reverse-transcription: 30 min at 60° C.; 2. PCR: 2 min at 94° C.,followed by 30-35 cycles (as optimized for each gene) for 30 s at 94°C., 30 s at optimized annealing temperature and 2 min at 68° C.; 3.final extension: 7 min at 68° C. PCR products were electrophoresed on1.5% agarose gels. Human (13-actin primers (5′-GCGAGAAGATGACCCAGATCAT-3′and 5′-GCTCAGGAGGAGCAATGATCTT-3′) were used as the internal control. TheRT-PCR analysis confirmed that the expression of the three secretedproteins: PBP, PF4 and F13A were all upregulated in CAD blood cells (seeFIG. 23). TABLE 5 Protein Accession Fold Functional Accession number(average) category Number Upregulated gene in CAD REV3-like, catalyticAF035537 2.3 Cell cycle NP_002903 subunit of DNA polymerase zetaTGFB1-induced anti- D86970 2.2 Cell cycle NP_510880 apoptotic factor 1 Adisintegrin and AA044656 2.7 Cell signaling NP_001101 metalloproteinasedomain 10 Centaurin, delta 2 AA351412 2 Cell signaling NP_631920Chloride intracellular AA411940 2.2 Cell signaling NP_039234 channel 4Endothelin receptor typeA D90348 2.1 Cell signaling NP_001948 Glutamatereceptor, N33821 2.4 Cell signaling NP_777567 ionotropicMitogen-activated protein L38486 3.7 Cell signaling NP_002395 kinase 7Mitogen-activated protein AB009356 4.5 Cell signaling NP_663306 kinasekinase kinase 7 Myristoylated alanine-rich D10522 2.5 Cell signalingNP_002347 protein kinase C substrate NIMA-related kinase 7 AA093324 3.5Cell signaling NP_598001 PAK2 AA262968 3.5 Cell signaling Q13177Phospholipid scramblase 1 AA054476 3.3 Cell signaling NP_066928 Serumdeprivation Z30112 4.5 Cell signaling NP_004648 response Adducin 3AA029158 2.9 Cell structure NP_063968 Desmin AF167579 4.4 Cell structureNP_001918 Fibromodulin W23613 2.9 Cell structure NP_002014 Laminin, beta2 S77512 2.2 Cell structure NP_002283 Laminin, beta 3 L25541 2.4 Cellstructure NP_000219 Osteonectin Y00755 3.1 Cell structure NP_003109 CD59antigen p18-20 W01111 2.4 Cell/organism NP_000602 defense ClusterinM64722 3.5 Cell/organism NP_001822 defense F13A M14539 2.1 Cell/organismNP_000120 defense Defensin, alpha 1 M26602 4.2 Cell/organism NP_004075defense PF4 M25897 2.1 Cell/organism NP_002610 defense PBP M54995 5.5Cell/organism NP_002695 defense E2F transcription factor 3 D38550 2.1Gene NP_001940 expression Early growth response 1 M62829 2.7 GeneNP_001955 expression Eukaryotic translation N86030 2.3 Gene NP_001393elongation factor 1 alpha 1 expression Eukaryotic translation M15353 2.1Gene NP_001959 initiation factor 4E expression F-box and WD-40 domainAB014596 2.7 Gene NP_387449 protein 1B expression Makorin, ring fingerAA331966 2.1 Gene NP_054879 protein, 2 expression Non-canonicalubiquitin- N92776 2.5 Gene NP_057420 conjugating enzyme 1 expressionNuclear receptor subfamily Z30425 4.7 Gene NP_005113 1, group I, member3 expression Ring finger protein 11 T08927 3 Gene NP_055187 expressionTransducin-like enhancer M99435 3.3 Gene NP_005068 of split 1 expressionAlkaline phosphatase, AB011406 2.2 Metabolism NP_000469liver/bone/kidney Annexin A3 M63310 3.4 Metabolism NP_005130 Branchedchain AA336265 4.8 Metabolism NP_005495.1 aminotransferase 1, cytosolicCytochrome b AF042500 2.5 Metabolism Glutaminase D30931 2.6 MetabolismNP_055720 Lysophospholipase I AF035293 2.8 Metabolism NP_006321 NADHdehydrogenase 1, AA056111 2.5 Metabolism NP_002485 subcomplex unknown 1,6 kDa Phosphofructokinase M26066 2.2 Metabolism NP_000280Ubiquinol-cytochrome c M22348 2.5 Metabolism NP_006285 reductase bindingprotein CGI-110 protein AA341061 2.4 Unclassified NP_057131 DactylidinH95397 2.7 Unclassified NP_112225 Deleted in split-hand/split- T245032.4 Unclassified NP_006295 foot 1 region Follistatin-like 1 R14219 2.7Unclassified NP_009016 FUS-interacting protein 1 W37945 2.8 UnclassifiedNP_473357 Hypothetical protein W47233 7 Unclassified NP_112201 FLJ12619Hypothetical protein from N68247 2.7 Unclassified EUROIMAGE 588495Hypothetical protein AA251423 2.2 Unclassified NP_057702 LOC51315KIAA1705 protein T80569 2.7 Unclassified NP_009121.1 Mesoderm inductionearly AI650409 2.2 Unclassified NP_065999 response 1 Phosphodiesterase4D- AA740661 2.5 Unclassified NP_055459 interacting proteinPreimplantation protein 3 D59087 2.5 Unclassified NP_056202 Putativenuclear protein W33098 2.8 Unclassified NP_115788 ORF1-FL49 Similar torat nuclear H09434 2.2 Unclassified Q9H1E3 ubiquitous casein kinase 2Similar to RIKEN AA297412 2.5 Unclassified T02670 Spectrin, betaAI334431 2.5 Unclassified Q01082 Stromal cell-derived factor H71558 4.1Unclassified NP_816929 receptor 1 Thioredoxin-related AA421549 2.8Unclassified NP_110437 protein Transmembrane 4 D29808 2.4 UnclassifiedNP_004606 superfamily member 2 Tumor endothelial marker 8 D79964 2.5Unclassified NP_444262 Downregulated gene in CAD CASP8 and FADD-likeAF015450 0.45 Cell cycle NP_003870 apoptosis regulator CD81 antigenM33680 0.41 Cell cycle NP_004347 Cell division cycle 25B M81934 0.4 Cellcycle NP_068660 DEAD/H (Asp-Glu-Ala- AA985699 0.42 Cell cycle NP_694705Asp/His) box polypeptide 27 F-box and leucine-rich R98291 0.27 Cellcycle NP_036440 repeat protein 11 Minichromosome H10286 0.43 Cell cycleNP_003897 maintenance deficient 3 associated protein Protein phosphatase2, J02902 0.48 Cell cycle NP_055040 regulatory subunit A, alpha isoformThyroid autoantigen 70 kDa J04607 0.25 Cell cycle NP_001460 Adisintegrin and R32760 0.37 Cell signaling metalloproteinase domain 17 Akinase anchor protein 13 M90360 0.31 Cell signaling NP_658913Calpastatin AF037194 0.39 Cell signaling NP_006471 Diacylglycerolkinase, AF064770 0.44 Cell signaling NP_001336 alpha 80 kDagamma-aminobutyric acid AJ012187 0.42 Cell signaling NP_068705 Breceptor, 1 Inositol polyphosphate-5- U84400 0.41 Cell signalingNP_005532 phosphatase, 145 kDa Lymphocyte-specific X05027 0.45 Cellsignaling NP_005347 protein tyrosine kinase RAP1B, member of RAS P095260.4 Cell signaling P09526 oncogene family Ras association AF061836 0.43Cell signaling NP_733835 (RalGDS/AF-6) domain family 1 CDC42-effectorprotein 3 AF104857 0.28 Cell signaling NP_006440 Leupaxin AF062075 0.31Cell signaling NP_004802 Annexin A6 D00510 0.45 Cell structure NP_004024RAN-binding protein 9 AB008515 0.41 Cell structure NP_005484 Thymosin,beta 10 M20259 0.26 Cell structure NP_066926 GranzymeA M18737 0.17Cell/organism NP_006135 defense ThromboxaneA synthase 1 M80646 0.44Cell/organism NP_112246 defense Coatomer protein AA357332 0.39 GeneNP_057535 complex, subunit beta expression Cold-inducible RNA- H398200.27 Gene NP_001271 binding protein expression Leucine-rich repeatU69609 0.44 Gene NP_004726 interacting protein 1 expression Proteasomesubunit, alpha D00762 0.31 Gene NP_687033 type, 3 expression Proteasomesubunit, alpha AF022815 0.35 Gene NP_689468 type, 7 expression Proteinphosphatase 1G, AI417405 0.5 Gene NP_817092 gamma isoform expressionRibonuclease/angiogenin M36717 0.44 Gene NP_002930 inhibitor expressionRNA-binding protein- AF021819 0.3 Gene NP_009193 regulatory subunitexpression Signal transducer and U16031 0.45 Gene NP_003144 activator oftranscription 6 expression Transcription factor A, M62810 0.41 GeneNP_036383 mitochondrial expression Ubiquitin-specific protease 4AF017306 0.31 Gene NP_003354 expression Dehydrogenase/reductase AA1000460.46 Metabolism NP_612461 SDR family member 1 Solute carrier family 25,J03592 0.3 Metabolism NP_001627 member 6 Amplified in osteosarcomaU41635 0.45 Unclassified NP_006803 Expressed in activated C00577 0.45Unclassified NP_009198 T/LAK lymphocytes Integral inner nuclear W004600.4 Unclassified NP_055134 membrane protein Phosphodiesterase 4D- T959690.45 Unclassified NP_055459 interacting protein Tumor endothelial markerN93789 0.45 Unclassified NP_065138 7 precursor Wiskott-Aldrich syndromeAF031588 0.22 Unclassified NP_003378 protein interacting protein

EXAMPLE 10

[0251] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andhypertension as compared with gene expression profiles from normalindividuals.

[0252] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith osteoarthritis and hypertension as compared to blood samples takenfrom healthy patients.

[0253] As used herein, the term “hypertension” is defined as high bloodpressure or elevated arterial pressure. Patients identified withhypertension herein include persons who have an increased risk ofdeveloping a morbid cardiovascular event and/or persons who benefit frommedical therapy designed to treat hypertension. Patients identified withhypertension also can include persons having systolic blood pressureof >130 mm Hg or a diastolic blood pressure of >90 mm Hg or a persontakes antihypertensive medication. Osteoarthritis (OA), as used hereinalso known as “degenerative joint disease”, represents failure of adiarthrodial (movable, synovial-lined) joint. It is a condition, whichaffects joint cartilage, and or subsequently underlying bone andsupporting tissues leading to pain, stiffness, movement problems andactivity limitations. It most often affects the hip, knee, foot, andhand, but can affect other joints as well.

[0254] OA severity can be graded according to the system described byMarshall (Marshall K W. J. Rheumatol, 1996:23(4) 582-85). Briefly, eachof the six knee articular surfaces was assigned a cartilage grade withpoints based on the worst lesion seen on each particular surface. Grade0 is normal (0 points), Grade I cartilage is soft or swollen but thearticular surface is intact (1 point). In Grade II lesions, thecartilage surface is not intact but the lesion does not extend down tosubchondral bone (2 points). Grade III damage extends to subchondralbone but the bone is neither eroded nor eburnated (3 points). In GradeIV lesions, there is ebumation of or erosion into bone (4 points). Aglobal OA score is calculated by summing the points from all sixcartilage surfaces. If there is any associated pathology, such asmeniscus tear, an extra point will be added to the global score. Basedon the total score, each patient is then categorized into one of four OAgroups: mild (1-6), moderate (7-12), marked (13-18), and severe (>18).As used herein, patients identified with OA may be categorized in any ofthe four OA groupings as described above.

[0255] Blood samples were taken from patients who were diagnosed withosteoarthritis and hypertension as defined herein. Gene expressionprofiles were then analysed and compared to profiles from patientsunaffected by any disease. In each case, the diagnosis of osteoarthritisand hypertension was corroborated by a skilled Board certifiedphysician.

[0256] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients with disease ascompared to healthy patients was determined by statistical analysisusing the Wilcox Mann Whitney rank sum test (Glantz S A., Primer ofBiostatistics., 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0257]FIG. 8 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having hypertension andosteoarthritis as compared with gene expression profiles from normalindividuals. Expression profiles were generated using GeneSpring™software analysis as described herein. Each column represents thehybridization pattern resulting from a single individual. In thisexample, hypertensive patients also presented with OA, as describedherein. Normal individuals have no known medical conditions and were nottaking any known medication. Hybridizations to create said geneexpression profiles were done using the ChondroChip™. A dendogramanalysis is shown above. Samples are clustered and marked asrepresenting patients who are hypertensive or normal. The “*” indicatesthose patients who abnormally clustered as either hypertensive, ornormal despite presenting with the reverse. The number of hybridizationsprofiles determined for either hypertensive patients or normalindividuals are shown. 861 differentially expressed genes wereidentified as being differentially expressed with a p value of <0.05 asbetween the hypertensive patients and normal individuals. The identityof the differentially expressed genes is shown in Table 3A.

[0258] Classification or class prediction of a test sample as eitherhaving hypertension and OA or being normal can be done using thedifferentially expressed genes as shown in Table 3A in combination withwell known statistical algorithms for class prediction as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 10A

[0259] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having osteoarthritis and hypertensionas compared with gene expression profiles from patients havingosteoarthritis only.

[0260] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken fromco-morbid patients with osteoarthritis and hypertension as compared toblood samples taken from OA patients only.

[0261] Blood samples were taken from patients who were diagnosed withosteoarthritis and hypertension as defined herein. Gene expressionprofiles were then analysed and compared to profiles from patientshaving OA only. In each case, the diagnosis of osteoarthritis and/orhypertension was corroborated by a skilled Board certified physician.

[0262] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients with disease ascompared to OA patients only was determined by statistical analysisusing the Wilcox Mann Whitney rank sum test (Glantz SA., Primer ofBiostatistics., 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0263] Expression profiles were generated using GeneSpring™ softwareanalysis as described herein (data not shown). The gene list generatedfrom this analysis was identified and those genes previously identifiedin Table 3A removed so as to identify those genes which are unique tohypertension. 790 differentially expressed genes were identified asbeing differentially expressed with a p value of <0.05 as between the OAand hypertensive patients when compared with OA individuals. 577 geneswere identified as unique to hypertension. The identity of thesedifferentially expressed genes are shown in Table 3P. A gene list isalso provided of the 213 genes which were found in common as betweenthose genes identified in Table 3A and genes differentially expressed inblood samples taken from patients with osteoarthritis and hypertensionas compared to blood samples taken from OA patients only. The identityof these intersecting differentially expressed genes is shown in Table3Q and a venn diagram showing the relationship between the variousgroups of gene lists is found in FIG. 29.

[0264] Classification or class prediction of a test sample as havinghypertension or not having hypertension can be done using thedifferentially expressed genes as shown in Table 3P as the predictorgenes in combination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.Classification of individuals as having both OA and hypertension usingthe genes in Table 3Q can also be performed.

EXAMPLE 11

[0265] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andobesity as compared with gene expression profiles from normalindividuals.

[0266] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith obesity and OA as compared to blood samples taken from healthypatients.

[0267] As used herein, “obesity” is defined as an excess of adiposetissue that imparts a health risk. Obesity is assessed in terms ofheight and weight in the relevance of age. Patients who are consideredobese include, but are not limited to, patients having a body mass indexor BMI ((defined as body weight in kg divided by (height in meters)²)greater than or equal to 30.0.

[0268] Blood samples were taken from patients who were diagnosed withosteoarthritis and obesity as defined herein. Gene expression profileswere then analysed and compared to profiles from patients unaffected byany disease. In each case, the diagnosis of the disease was corroboratedby a skilled Board certified physician. Total mRNA from a drop ofperipheral whole blood taken from each patient was isolated usingTRIzol® reagent (GIBCO) and fluorescently labelled probes for each bloodsample were generated as described above. Each probe was denatured andhybridized to a 15K Chondrogene Microarray Chip (ChondroChip™) asdescribed herein. Identification of genes differentially expressed inblood samples from patients with disease as compared to healthy patientswas determined by statistical analysis using the Wilcox Mann Whitneyrank sum test (Glantz S A., Primer of Biostatistics., 5th ed., New York,USA: McGraw-Hill Medical Publishing Division, 2002).

[0269]FIG. 9 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as obeseas described herein as compared with gene expression profiles fromnormal individuals. Expression profiles were generated using GeneSpring™software analysis as described herein. Each column represents thehybridization pattern resulting from a single individual. In thisexample, obese patients also presented with OA, as described herein.Normal individuals have no known medical conditions and were not takingany known medication. Hybridizations to create said gene expressionprofiles were done using the ChondroChip™. A dendogram analysis is shownabove. Samples are clustered and marked as representing patients who areobese or normal. The “*” indicates those patients who abnormallyclustered as either obese or normal despite presenting with the reverse.The number of hybridization profiles determined for obese patients withOA and normal individuals are shown. 913 genes were identified as beingdifferentially expressed with a p value of <0.05 as between the obesepatients with OA and normal individuals is noted. The identity of thedifferentially expressed genes is shown in Table 3B.

[0270] Classification or class prediction of a test sample as eitherhaving obesity and OA or being normal can be done using thedifferentially expressed genes as shown in Table 3B in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 11A

[0271] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andobesity as compared with gene expression profiles from patients havingosteoarthritis only.

[0272] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith obesity and OA as compared to blood samples taken from patientswith OA only.

[0273] Blood samples were taken from patients who were diagnosed withosteoarthritis and obesity as defined herein. Gene expression profileswere then analysed and compared to profiles from patients affected by OAonly.

[0274] In each case, the diagnosis of the disease was corroborated by askilled Board certified physician. Total mRNA from a drop of peripheralwhole blood taken from each patient was isolated using TRIzol® reagent(GIBCO) and fluorescently labelled probes for each blood sample weregenerated as described above. Each probe was denatured and hybridized toa 15K Chondrogene Microarray Chip (ChondroChip™) as described herein.Identification of genes differentially expressed in blood samples frompatients with obesity and OA as compared to OA patients only wasdetermined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A., Primer of Biostatistics., 5th ed. New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0275] Expression profiles were generated using GeneSpring™ softwareanalysis as described herein (data not shown). 671 genes were identifiedas being differentially expressed with a p value of <0.05 as between theobese patients with OA and those patients with only OA. Those genespreviously identified in Table 3B were removed so as to identify thosegenes which are unique to obesity. The identity of these 519 genesunique to obesity are shown in Table 3R. A gene list is also provided ofthose genes which were found in common as between those genes identifiedin Table 3B and genes differentially expressed in blood samples takenfrom patients with osteoarthritis and obesity as compared to bloodsamples taken from OA patients only. 152 genes are shown in Table 3S. Avenn diagram showing the relationship between the various groups of genelists is found in FIG. 30.

[0276] Classification or class prediction of a test sample as havingobesity or not having obesity can be done using the differentiallyexpressed genes as shown in Table 3R as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.Classification of individuals as having both OA and obesity using thegenes in Table 3S can also be performed.

EXAMPLE 12

[0277] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andallergies as compared with gene expression profiles from normalindividuals.

[0278] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith allergies as compared to blood samples taken from healthy patients.

[0279] As used herein, “allergies” encompasses diseases and conditionswherein a patient demonstrates a hypersensitive or allergic reaction toone or more substances or stimuli such as drugs, food stuffs, plants,animals etc. and as a result has an increased immune response. Suchimmune responses can include anaphylaxis, allergic rhinitis, asthma,skin sensitivity such as urticaria, eczema, and allergic contactdermatitis and ocular allergies such as allergic conjunctivitis andcontact allergy. Patients identified as having allergies includespatients having one or more of the above noted conditions.

[0280] Blood samples were taken from patients who were diagnosed withosteoarthritis and allergies as defined herein. These patients areclassified as presenting with co-morbidity, or multiple disease states.Gene expression profiles were then analysed and compared to profilesfrom patients unaffected by any disease. In each case, the diagnosis ofosteoarthritis and allergies was corroborated by a skilled Boardcertified physician.

[0281] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients withosteoarthritis and allergies as compared to healthy patients wasdetermined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A., Primer of Biostatistics., 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0282]FIG. 10 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingallergies as described herein as compared with gene expression profilesfrom normal individuals. Expression profiles were generated usingGeneSpring™ software analysis as described herein. Each columnrepresents the hybridization pattern resulting from a single individual.In this example, patients with allergies also presented with OA, asdescribed herein. Normal individuals had no known medical conditions andwere not taking any known medication. Hybridizations to create said geneexpression profiles were done using the ChondroChip™. A dendogramanalysis is shown above. Samples are clustered and marked asrepresenting patients who are obese or normal. The “*” indicates thosepatients who abnormally clustered as either having allergies or beingnormal despite presenting with the reverse. The number of hybridizationsprofiles determined for patients with allergies and normal individualsare shown. 633 genes were identified as being differentially expressedwith a p value of <0.05 as between patients with allergies and normalindividuals is noted. The identity of the differentially expressed genesis shown in Table 3C.

[0283] Classification or class prediction of a test sample as eitherhaving allergies and OA or being normal can be done using thedifferentially expressed genes as shown in Table 3C in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 12A

[0284] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having osteoarthritis (OA) andallergies as compared with gene expression profiles from individualswith OA only.

[0285] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith allergies and OA as compared to blood samples taken from OApatients.

[0286] Blood samples were taken from patients who were diagnosed withosteoarthritis and allergies as defined herein. Gene expression profileswere then analysed and compared to profiles from patients affected by OAonly. In each case, the diagnosis of osteoarthritis and allergies wascorroborated by a skilled Board certified physician.

[0287] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients withosteoarthritis and allergies as compared to OA patients only wasdetermined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A., Primer of Biostatistics., 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0288] Expression profiles were generated using GeneSpring™ softwareanalysis as described herein (data not shown). 498 genes were identifiedas being differentially expressed with a p value of <0.05 as betweenpatients with allergies and OA as compared with patients with OA only.Of the 498 genes identified, those genes previously identified in Table3C were removed so as to identify those genes which are unique toallergies. 257 differentially expressed genes were identified as beingas unique to allergies. The identity of these differentially expressedgenes is shown in Table 3T. A gene list is also provided of the 241genes which were found in common as between those genes identified inTable 3C and genes differentially expressed in blood samples taken frompatients with osteoarthritis and allergies as compared to blood samplestaken from OA patients only. The identity of these intersectingdifferentially expressed genes is shown in Table 3U and a venn diagramshowing the relationship between the various groups of gene lists isfound in FIG. 31.

[0289] Classification or class prediction of a test sample as havingallergies or not having allergies can be done using the differentiallyexpressed genes as shown in Table 3T as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.Classification of individuals as having both OA and allergies using thegenes in Table 3U can also be performed.

EXAMPLE 13

[0290] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andsubject to systemic steroids as compared with gene expression profilesfrom normal individuals

[0291] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientssubject to systemic steroids as compared to blood samples taken fromhealthy patients.

[0292] As used herein, “systemic steroids” indicates a person subjectedto artificial levels of steroids as a result of medical intervention.Such systemic steroids include birth control pills, prednisone, andhormones as a result of hormone replacement treatment. A personidentified as having systemic steroids is one who is on one or more ofthe following of the above treatment regimes.

[0293] Blood samples were taken from patients who were diagnosed withosteoarthritis and subject to systemic steroids as defined herein. Geneexpression profiles were then analysed and compared to profiles frompatients unaffected by any disease. In each case, the diagnosis ofosteoarthritis and systemic steroids was corroborated by a skilled Boardcertified physician.

[0294] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to the 15K ChondrogeneMicroarray Chip (ChondroChip™) as described herein. Identification ofgenes differentially expressed in blood samples from patients withosteoarthritis and subject to systemic steroids as compared to healthypatients was determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A., Primer of Biostatistics., 5th ed.,New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0295]FIG. 11 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were subject to systemicsteroids as described herein as compared with gene expression profilesfrom normal individuals. Expression profiles were generated usingGeneSpring™ software analysis as described herein. Each columnrepresents the hybridization pattern resulting from a single individual.In this example, patients taking systemic steroids also presented withOA, as described herein. Normal individuals have no known medicalconditions and were not taking any known medication. Hybridizations tocreate said gene expression profiles were done using the ChondroChip™.(A dendogram analysis is shown above. Samples are clustered and markedas representing patients who are taking systemic steroids or normal. The“*” indicates those patients who abnormally clustered as either systemicsteroids or normal despite presenting with the reverse. The number ofhybridizations profiles determined for patients with systemic steroidsand normal individuals are shown. 605 genes were identified as beingdifferentially expressed with a p value of <0.05 as-between patientswith systemic steroids and normal individuals is noted. The identity ofthe differentially expressed genes is shown in Table 3D.

[0296] Classification or class prediction of a test sample from apatient as indicating said patient takes systemic steroids and has OA oras being normal can be done using the differentially expressed genes asshown in Table 3A in combination with well known statistical algorithmsfor class prediction as would be understood by a person skilled in theart and is described herein. Commercially available programs such asthose provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 13A

[0297] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andsubject to systemic steroids as compared with gene expression profilesfrom with osteoarthritis only.

[0298] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientssubject to systemic steroids and having OA as compared to blood samplestaken from OA patients only.

[0299] Blood samples were taken from patients who were diagnosed withosteoarthritis and subject to systemic steroids as defined herein. Geneexpression profiles were then analysed and compared to profiles frompatients having OA only. In each case, the diagnosis of osteoarthritisand systemic steroids was corroborated by a skilled Board certifiedphysician.

[0300] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to the 15K ChondrogeneMicroarray Chip (ChondroChip™) as described herein. Identification ofgenes differentially expressed in blood samples from patients withosteoarthritis and subject to systemic steroids as compared patientswith OA only was determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A., Primer of Biostatistics., 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0301] Expression profiles were generated using GeneSpring™ softwareanalysis as described herein (data not shown). 553 genes were identifiedas being differentially expressed with a p value of <0.05 as betweenpatients taking systemic steroids and OA as compared with patients withOA only. Of the 553 genes identified, those genes previously identifiedin Table 3D were removed so as to identify those genes which are uniqueto systemic steroids. 362 differentially expressed genes were identifiedas being as unique to systemic steroids. The identity of thesedifferentially expressed genes are shown in Table 3V. A gene list isalso provided of the 191 genes which were found in common as betweenthose genes identified in Table 3D and genes differentially expressed inblood samples taken from patients with osteoarthritis and systemicsteroids as compared to blood samples taken from OA patients only. Theidentity of these intersecting differentially expressed genes is shownin Table 3W and a venn diagram showing the relationship between thevarious groups of gene lists is found in FIG. 32.

[0302] Classification or class prediction of a test sample of anindividual as either taking systemic steroids or not taking systemicsteroids can be done using the differentially expressed genes as shownin Table 3V as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics (e.g. GeneSpring™) for Class Predicationare also available. Classification of individuals as having both OA andtaking systemic steroids using the genes in Table 3W can also beperformed.

EXAMPLE 13B

[0303] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from co-morbid individuals having osteoarthritis andsubject to systemic steroids as compared with gene expression profilesfrom normal individuals.

[0304] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientssubject to various specific systemic steroids as compared to bloodsamples taken from healthy patients, and the ability to categorize anddifferentiate as between the systemic steroid being taken.

[0305] As used herein, “systemic steroids” indicates a person subjectedto artificial levels of steroids as a result of medical intervention.Such systemic steroids include birth control pills, prednisone, andhormones as a result of hormone replacement treatment. A personidentified as having systemic steroids is one who is on one or more ofthe following of the above treatment regimes.

[0306] Blood samples were taken from patients who were diagnosed withosteoarthritis and subject to systemic steroids as defined herein. Geneexpression profiles were then analysed and compared as between thesystemic steroids as compared to profiles from patients unaffected byany disease. In each case, the diagnosis of osteoarthritis and systemicsteroids was corroborated by a skilled Board certified physician.

[0307] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to the 15K ChondrogeneMicroarray Chip (ChondroChip™) as described herein. Identification ofgenes differentially expressed in blood samples from patients withosteoarthritis and subject to systemic steroids as compared to healthypatients was determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A., Primer of Biostatistics., 5th ed.,New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0308]FIG. 34 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were subject to eitherbirth control, prednisone, or hormone replacement therapy as describedherein as compared with gene expression profiles from normalindividuals. Expression profiles were generated using GeneSpring™software analysis as described herein. Each column represents thehybridization pattern resulting from a single individual. In thisexample, patients taking with each of the systemic steroids alsopresented with OA, as described herein. Normal individuals have no knownmedical conditions and were not taking any known medication.Hybridizations to create said gene expression profiles were done usingthe ChondroChip™. A dendogram analysis is shown above. Samples areclustered and marked as representing patients who are taking birthcontrol, prednisone, hormone replacement therapy or normal. The “*”indicates those patients who abnormally clustered. The number ofhybridizations profiles determined for patients with birth control,prednisone, hormone replacement therapy or normal individuals are shown.396 genes were identified as being differentially expressed with a pvalue of <0.05 as between patients with systemic steroids and normalindividuals is noted. The identity of the differentially expressed genesis shown in Table 3AD.

[0309] Classification or class prediction of a test sample from apatient as indicating said patient takes systemic steroids and has OA oras being normal can be done using the differentially expressed genes asshown in Table 3AD in combination with well known statistical algorithmsfor class prediction as would be understood by a person skilled in theart and is described herein. Commercially available programs such asthose provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 14

[0310] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having hypertension as compared withgene expression profiles from normal individuals.

[0311] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith hypertension but without osteoarthritis as compared to bloodsamples taken from healthy patients.

[0312] As used herein, the term “hypertension” is defined as high bloodpressure or elevated arterial pressure. Patients identified withhypertension herein include persons who have an increased risk ofdeveloping a morbid cardiovascular event and/or persons who benefit frommedical therapy designed to treat hypertension. Patients identified withhypertension also can include persons having systolic blood pressureof >130 mm Hg or a diastolic blood pressure of >90 mm Hg or a persontakes antihypertensive medication.

[0313] Blood samples were taken from patients who were diagnosed withhypertension as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of hypertension was corroborated bya skilled Board certified physician.

[0314] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients withhypertension as compared to healthy patients was determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A., Primer of Biostatistics., 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0315]FIG. 12 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having hypertension ascompared with gene expression profiles from samples of bothnon-hypertensive and normal individuals. Expression profiles weregenerated using GeneSpring™ software analysis as described herein. Eachcolumn represents the hybridization pattern resulting from a singleindividual. Non-hypertensive individuals presented without hypertension,but may have presented with other medical conditions and may be undervarious treatment regimes. Normal individuals have no known medicalconditions and were not taking any known medication. Hybridizations tocreate said gene expression profiles were done using the ChondroChip™. Adendogram analysis is shown above. Samples are clustered and marked asrepresenting patients who are hypertensive, normal or non-hypertensive.The “*” indicates those patients who abnormally clustered as eitherhypertensive, non-hypertensive or normal despite actual presentation.The number of hybridizations profiles determined for hypertensivepatients, non-hypertensive patients and normal individuals are shown.1,993 genes identified as being differentially expressed with a p valueof <0.05 as between the hypertensive patients and the combined normaland non-hypertensive individuals is noted. The identity of thedifferentially expressed genes are shown in Table 3E.

[0316] Classification or class prediction of a test sample of anindividual so as to determine whether said individual has or does nothave hypertension can be done using the differentially expressed genesas shown in Table 3E as the predictor genes in combination with wellknown statistical algorithms as would be understood by a person skilledin the art and described herein. Commercially available programs such asthose provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 15

[0317] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having obesity as compared with geneexpression profiles from normal individuals.

[0318] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith obesity but without osteoarthritis as compared to blood samplestaken from healthy patients.

[0319] As used herein, “obesity” is defined as an excess of adiposetissue that imparts a health risk. Obesity is assessed in terms ofheight and weight in the relevance of age. Patients who are consideredobese include, but are not limited to, patients having a body mass indexor BMI ((defined as body weight in kg divided by (height in meters)²)greater than or equal to 30.0.

[0320] Blood samples were taken from patients who were diagnosed withhypertension as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of obesity was corroborated by askilled Board certified physician.

[0321] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a (ChondroChip™) as describedherein. Identification of genes differentially expressed in bloodsamples from patients with obesity as compared to healthy patients wasdetermined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A., Primer of Biostatistics, 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0322]FIG. 13 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as obeseas described herein as compared with gene expression profiles fromnormal and non-obese individuals. Expression profiles were generatedusing GeneSpring™ software analysis as described herein. Each columnrepresents the hybridization pattern resulting from a single individual.Normal individuals have no known medical conditions and were not takingany known medication. Non-obese individuals presented without obesity,but may have presented with other medical conditions and may be undervarious treatment regimes. Hybridizations to create said gene expressionprofiles were done using the ChondroChip™. A dendograrm analysis isshown above. Samples are clustered and marked as representing patientswho are obese, normal or non-obese. The “*” indicates those patients whoabnormally clustered as either obese, normal or non-obese despite actualpresentation. The number of hybridizations profiles determined for obesepatients, non-obese patients and normal individuals are shown. 1,147genes were identified as being differentially expressed with a p valueof <0.05 as between the obese patients and the combination of normal andnon-obese individuals is noted. The identity of the differentiallyexpressed genes is shown in Table 3F.

[0323] Classification or class prediction of a test sample as beingobese or not being obese can be done using the differentially expressedgenes as shown in Table 3F as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 16

[0324] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having type 2 diabetes as comparedwith gene expression profiles from normal individuals.

[0325] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith type 2 diabetes but without osteoarthritis as compared to bloodsamples taken from healthy patients.

[0326] As used herein, “diabetes”, or “diabetes mellitus” includes both“type 1 diabetes” (insulin-dependent diabetes (IDDM)) and “type 2diabetes” (insulin-independent diabetes (NIDDM). Both type 1 and type 2diabetes characterized in accordance with Harrison's Principles ofInternal Medicine 14th edition, as a person having a venous plasmaglucose concentration>140 mg/dL on at least two separate occasions afterovernight fasting and venous plasma glucose concentration>200 mg/dL at 2h and on at least one other occasion during the 2-h test followingingestion of 75g of glucose. Patients identified as having type 2diabetes as described herein are those demonstrating insulin-independentdiabetes as determined by the methods described above.

[0327] Blood samples were taken from patients who were diagnosed withtype II diabetes as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of type II diabetes wascorroborated by a skilled Board certified physician.

[0328] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients with type 2diabetes as compared to healthy patients was determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A.,Primer of Biostatistics., 5th ed., New York, USA: McGraw-Hill MedicalPublishing Division, 2002).

[0329]FIG. 14 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingtype 2 diabetes as described herein as compared with gene expressionprofiles from normal and non-type 2 diabetes individuals. Expressionprofiles were generated using GeneSpring™ software analysis as describedherein. Each column represents the hybridization pattern resulting froma single individual. Normal individuals have no known medical conditionsand were not taking any known medication. Non-type 2 diabetesindividuals presented without type 2 diabetes, but may have presentedwith other medical conditions and may be under various treatmentregimes. Hybridizations to create said gene expression profiles weredone using the ChondroChip™. A dendogram analysis is shown above.Samples are clustered and marked as representing patients who have type2 diabetes, are normal or do not have type 2 diabetes. The “*” indicatesthose patients who abnormally clustered despite actual presentation. Thenumber of hybridizations profiles determined for type 2 diabetes,non-type 2 diabetes and normal individuals are shown. 915 wereidentified as being differentially expressed with a p value of <0.05 asbetween the type 2 diabetes patients and the combination of normal andnon type 2 diabetes individuals is noted. The identity of thedifferentially expressed genes is shown in Table 3G.

[0330] Classification or class prediction of a test sample of anindividual so as to determine whether said individual has type 2diabetes or does not have type 2 diabetes can be done using thedifferentially expressed genes as shown in Table 3G as the predictorgenes in combination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 17

[0331] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having hyperlipidemia as compared withgene expression profiles from normal individuals.

[0332] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith hyperlipidemia but without osteoarthritis as compared to bloodsamples taken from healthy patients.

[0333] As used herein, “hyperlipidemia” is defined as an elevation oflipid protein profiles and includes the elevation of chylomicrons, verylow-density lipoproteins (VLDL), intermediate-density lipoproteins(IDL), low-density lipoproteins (LDL), and/or high-density lipoproteins(HDL) as compared with the general population. Hyperlipidemia includeshypercholesterolemia and/or hypertriglyceridemia. Byhypercholesterolemia, it is meant elevated fasting plasma totalcholesterol level of >200 mg/dL, and/or LDL-cholesterol levels of >130mg/dL. A desirable level of HDL-cholesterol is >60 mg/dL. Byhypertriglyceridemia it is meant plasma triglyceride (TG) concentrationsof greater than the 90^(th) or 95^(th) percentile for age and sex andcan include, for example, TG>160 mg/dL as determined after an overnightfast.

[0334] Blood samples were taken from patients who were diagnosed withhyperlipidemia as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of hyperlipidemia was corroboratedby a skilled Board certified physician.

[0335] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients withhyperlipidemia as compared to healthy patients was determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A., Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0336]FIG. 15 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havinghyperlipidemia as described herein as compared with gene expressionprofiles from normal and non-hyperlipidemia patients. Expressionprofiles were generated using GeneSpring™ software analysis as describedherein. Each column represents the hybridization pattern resulting froma single individual. Normal individuals have no known medical conditionsand were not taking any known medication. Non hyperlipidemia individualspresented without elevated cholesterol or elevated triglycerides but mayhave presented with other medical conditions and may be under varioustreatment regimes. Hybridizations to create said gene expressionprofiles were done using the ChondroChip™. A dendogram analysis is shownabove. Samples are clustered and marked as representing patients whohave elevated lipids and/or cholesterol, are normal or do not haveelevated lipids or cholesterol. The “*” indicates those patients whoabnormally clustered as having either hyperlipidemia, normal ornon-hyperlipidemia despite actual presentation. The number ofhybridizations profiles determined for hyperlipidemia patients,non-hyperlipidemia patients and normal individuals are shown. 1,022genes were identified as being differentially expressed with a p valueof <0.05 as between the patients with hyperlipidemia and the combinationof normal and non hyperlipidemia individuals. The identity of thedifferentially expressed genes is shown in Table 3H.

[0337] Classification or class prediction of a test sample of anindividual as having hyperlipidemia or not having hyperlipidemia can bedone using the differentially expressed genes as shown in Table 3H asthe predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics for Class Predication (e.g. GeneSpring™) are alsoavailable.

EXAMPLE 18

[0338] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having lung disease as compared withgene expression profiles from normal individuals.

[0339] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith lung disease but without osteoarthritis as compared to bloodsamples taken from healthy patients.

[0340] As used herein, “lung disease” encompasses any disease thataffects the respiratory system and includes bronchitis, chronicobstructive lung disease, emphysema, asthma, and lung cancer. Patientsidentified as having lung disease includes patients having one or moreof the above noted conditions.

[0341] Blood samples were taken from patients who were diagnosed withlung disease as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of lung disease was corroborated bya skilled Board certified physician.

[0342] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients with lungdisease as compared to healthy patients was determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A.,Primer of Biostatistics., 5th ed., New York, USA: McGraw-Hill MedicalPublishing Division, 2002).

[0343]FIG. 16 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havinglung disease as described herein as compared with gene expressionprofiles from normal and non lung disease individuals. Expressionprofiles were generated using GeneSpring™ software analysis as describedherein. Each column represents the hybridization pattern resulting froma single individual. Normal individuals have no known medical conditionsand were not taking any known medication. Non-lung disease individualspresented without lung disease, but may have presented with othermedical conditions and may be under various treatment regimes.Hybridizations to create said gene expression profiles were done usingthe ChondroChip™. A dendogram analysis is shown above. Samples areclustered and marked as representing patients who have lung disease, arenormal or do not have lung disease. The “*” indicates those patients whoabnormally clustered despite actual presentation. The number ofhybridizations profiles determined for either the lung disease patients,non-lung disease patients and normal individuals are show. 596 geneswere identified as being differentially expressed with a p value of<0.05 as between the lung disease patients and the combination of normaland non lung disease individuals is noted. The identity of thedifferentially expressed genes is shown in Table 31.

[0344] Classification or class prediction of a test sample of anindividual to determine whether said individual has lung disease or doesnot having lung disease can be done using the differentially expressedgenes as shown in Table 31 as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 19

[0345] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having bladder cancer ascompared with gene expression profiles from normal individuals.

[0346] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith bladder cancer but without osteoarthritis as compared to bloodsamples taken from healthy patients.

[0347] As used herein, the term “cancer” or “carcinoma” is defined as adisease in which cells behave abnormally and includes; (i) cancers whichoriginate from a single cell proliferating to form a clone of malignantcells, (ii) cancers wherein the growth of the cell is not regulated bynormal biological and physical influences of the environment, (iii)anaplasic cancer, wherein the cells lack normal coordinated celldifferentiation and (iv) metastasis cancer, wherein the cells have thecapacity for discontinuous growth and dissemination to other parts ofthe body. The diagnosis of cancer can include careful clinicalassessment and/or diagnostic investigations including endoscopy,imaging, histopathology, cytology and laboratory studies.

[0348] As used herein, “bladder cancer” includes carcinomas that occurin the transitional epithelium lining the urinary tract, starting at therenal pelvis and extending through the ureter, the urinary bladder, andthe proximal two-thirds of the urethra. As used herein, patientsdiagnosed with bladder cancer include patients diagnosed utilizing anyof the following methods or a combination thereof: urinary cytologicevaluation, endoscopic evaluation for the presence of malignant cells,CT (computed tomography), MRI (magnetic resonance imaging) formetastasis status.

[0349] Blood samples were taken from patients who were diagnosed withbladder cancer as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of bladder cancer was corroboratedby a skilled Board certified physician.

[0350] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with bladder cancer as compared to healthypatients was determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A., Primer of Biostatistics., 5th ed.,New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0351]FIG. 17 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingbladder cancer as described herein as compared with gene expressionprofiles from non bladder cancer individuals. Expression profiles weregenerated using GeneSpring™ software analysis as described herein. Eachcolumn represents the hybridization pattern resulting from a singleindividual. Non bladder cancer individuals presented without bladdercancer, but may have presented with other medical conditions and may beunder various treatment regimes. Hybridizations to create said geneexpression profiles were done using the Affymetrix U133A chip. Adendogram analysis is shown above. Samples are clustered and marked asrepresenting patients who have bladder cancer, or do not have bladdercancer. The “*” indicates those patients who abnormally clustered aseither bladder cancer, or non bladder cancer despite actualpresentation. The number of hybridizations profiles determined forpatients with bladder cancer and without bladder cancer are shown. 4,228genes were identified as being differentially expressed with a p valueof <0.05 as between the bladder cancer patients and the non bladdercancer individuals is noted. The identity of the differentiallyexpressed genes is shown in Table 3J.

[0352] Classification or class prediction of a test sample of anindividual to determine whether said individual has bladder cancer ordoes not having bladder cancer can be done using the differentiallyexpressed genes as shown in Table 3J as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 20

[0353] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having early or advancedbladder cancer as compared with gene expression profiles from normalindividuals.

[0354] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith early or advanced late stage bladder cancer but withoutosteoarthritis as compared to blood samples taken from healthy patients.

[0355] As used herein, “early stage bladder cancer” includes bladdercancer wherein the detection of the anatomic extent of the tumour, bothin its primary location and in metastatic sites, as defined by the TNMstaging system in accordance with Harrison's Principles of InternalMedicine 14th edition can be considered early stage. More specifically,early stage bladder cancer can include those instances wherein thecarcinoma is mainly superficial.

[0356] As used herein, “advanced stage bladder cancer” is defined asbladder cancer wherein the detection of the anatomic extent of thetumour, both in its primary location and in metastatic sites, as definedby the TNM staging system in accordance with Harrison's Principles ofInternal Medicine 14th edition, can be considered as advanced stage.More specifically, advanced stage carcinomas can involve instanceswherein the cancer has infiltrated the muscle and wherein metastasis hasoccurred.

[0357] Blood samples were taken from patients who were diagnosed withearly or advanced late stage bladder cancer as defined herein. Geneexpression profiles were then analysed and compared to profiles frompatients unaffected by any disease. In each case, the diagnosis of earlyor advanced late stage bladder cancer was corroborated by a skilledBoard certified physician.

[0358] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with early or advanced late stage bladdercancer as compared to healthy patients was determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A.,Primer of Biostatistics., 5th ed., New York, USA: McGraw-Hill MedicalPublishing Division, 2002).

[0359]FIG. 18 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingadvanced stage bladder cancer or early stage bladder cancer as describedherein as compared with gene expression profiles from non bladder cancerindividuals. Expression profiles were generated using GeneSpring™software analysis as described herein. Each column represents thehybridization pattern resulting from a single individual. Non bladdercancer individuals presented without bladder cancer, but may havepresented with other medical conditions and may be under varioustreatment regimes. Hybridizations to create said gene expressionprofiles were done using the Affymetrix U1338 chip. A dendogram analysisis shown above. Samples are clustered and marked as representingpatients who have early stage bladder cancer, advanced stage bladdercancer, or do not have bladder cancer. The “*” indicates those patientswho abnormally clustered despite actual presentation. The number ofhybridizations profiles determined for either early stage bladdercancer, advanced bladder cancer or non-bladder cancer are shown. 3,518genes were identified as being differentially expressed with a p valueof <0.05 as between the bladder cancer patients and the non bladdercancer individuals is noted. The identity of the differentiallyexpressed genes is shown in Table 3K.

[0360] Classification or class prediction of a test sample of anindividual to determine whether said individual has advanced bladdercancer, early stage bladder cancer or does not have bladder cancer canbe done using the differentially expressed genes as shown in Table 3K asthe predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 21

[0361] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having coronary arterydisease as compared with gene expression profiles from normalindividuals.

[0362] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith coronary artery disease but without osteoarthritis as compared toblood samples taken from healthy patients

[0363] As used herein, “Coronary artery disease” (CAD) is defined as acondition wherein at least one coronary artery has >50% luminal diameterstenosis, as diagnosed by coronary angiography and includes conditionsin which there is atheromatous narrowing and subsequent occlusion of thevessel. CAD includes those conditions which manifest as angina, silentischaemia, unstable angina, myocardial infarction, arrhythmias, heartfailure, and sudden death. Patients identified as having CAD hereinCoronary artery disease is defined

[0364] Blood samples were taken from patients who were diagnosed withCoronary artery disease as defined herein. Gene expression profiles werethen analysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of Coronary artery disease wascorroborated by a skilled Board certified physician.

[0365] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with Coronary artery disease as compared tohealthy patients was determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A., Primer of Biostatistics, 5thed., New York, USA, McGraw-Hill Medical Publishing Division, 2002).

[0366]FIG. 19 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingcoronary artery disease (CAD) as described herein as compared with geneexpression profiles from non-coronary artery disease individuals.Expression profiles were generated using GeneSpring™ software analysisas described herein. Each column represents the hybridization patternresulting from a single individual. Non coronary artery diseaseindividuals presented without coronary artery disease, but may havepresented with other medical conditions and may be under varioustreatment regimes. Hybridizations to create said gene expressionprofiles were done using the Affymetrix™ U133A chip. A dendogramanalysis is shown above. Samples are clustered and marked asrepresenting patients who have coronary artery disease or do not havecoronary artery disease. The “*” indicates those patients who abnormallyclustered despite actual presentation. The number of hybridizationsprofiles determined for patients with CAD or without CAD are shown. 967genes were identified as being differentially expressed with a p valueof <0.05 as between the coronary artery disease patients and thoseindividuals without coronary artery disease is noted. The identity ofthe differentially expressed genes is shown in Table 3L.

[0367] Classification or class prediction of a test sample of anindividual to determine whether said individual has CAD or does not haveCAD can be done using the differentially expressed genes as shown inTable 3L as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics for Class Predication (e.g. GeneSpring™)are also available.

EXAMPLE 22

[0368] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having Rheumatoid arthritisas compared with gene expression profiles from normal individuals.

[0369] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith Rheumatoid arthritis but without osteoarthritis as compared toblood samples taken from healthy patients.

[0370] Rheumatoid arthritis (RA) is defined as a chronic, multisystemdisease of unknown etiology with the characteristic feature ofpersistent inflammatory synovitis. Said inflammatory synovitis usuallyinvolves peripheral joints in a systemic distribution. Patients havingRA as defined herein were identified as having one or more of thefollowing; (i) cartilage destruction, (ii) bone erosions, and/or (iii)joint deformities.

[0371] Blood samples were taken from patients who were diagnosedRheumatoid arthritis as defined herein. Gene expression profiles werethen analysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of Rheumatoid arthritis wascorroborated by a skilled Board certified physician.

[0372] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with Rheumatoid arthritis as compared tohealthy patients was determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A., Primer of Biostatistics., 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0373]FIG. 20 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingrheumatoid arthritis as described herein as compared with geneexpression profiles from non-rheumatoid arthritis individuals.Expression profiles were generated using GeneSpring™ software analysisas described herein. Each column represents the hybridization patternresulting from a single individual. Normal individuals have no knownmedical conditions and were not taking any known medication. Nonrheumatoid arthritis individuals presented without rheumatoid arthritis,but may have presented with other medical conditions and may be undervarious treatment regimes. Hybridizations to create said gene expressionprofiles were done using ChondroChip™. A dendogram analysis is shownabove. Samples are clustered and marked as representing patients whohave rheumatoid arthritis or do not have rheumatoid arthritis. The “*”indicates those patients who abnormally clustered despite actualpresentation. The number of hybridizations profiles determined forpatients with rheumatoid arthritis and without rheumatoid arthritis areshown. 2,068 genes were identified as being differentially expressedwith a p value of <0.05 as between the rheumatoid arthritis patients anda combination of those individuals without rheumatoid arthritis andnormal is noted. The identity of the differentially expressed genes isshown in Table 3M.

[0374] Classification or class prediction of a test sample of anindividual as having rheumatoid arthritis or not having rheumatoidarthritis can be done using the differentially expressed genes as shownin Table 3M as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics for Class Predication (e.g. GeneSpring™)are also available.

EXAMPLE 23

[0375] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having depression as comparedwith gene expression profiles from normal individuals.

[0376] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith depression but without osteoarthritis as compared to blood samplestaken from healthy patients

[0377] As used herein “mood disorders” are conditions characterized by adisturbance in the regulation of mood, behavior, and affect. “Mooddisorders” can include depression, anxiety, schizophrenia, bipolardisorder, manic depression and the like.

[0378] As used herein “depression” includes depressive disorders ordepression in association with medical illness or substance abuse inaddition to depression as a result of sociological situations. Patientsdefined as having depression were diagnosed mainly on the basis ofclinical symptoms including a depressed mood episode wherein a persondisplays a depressed mood on a daily basis for a period of greater than2 weeks. A depressed mood episode may be characterized by sadness,indifference, apathy, or irritability and is usually associated withchanges in a number of neurovegetative functions, including sleeppatterns, appetite and weight, fatigue, impairment in concentration anddecision making.

[0379] Blood samples were taken from patients who were diagnosed withdepression as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of depression was corroborated by askilled Board certified physician.

[0380] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with depression as compared to healthypatients was determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A., Primer of Biostatistics, 5th ed.,New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0381]FIG. 21 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingdepression as described herein as compared with gene expression profilesfrom non-depression individuals. Expression profiles were generatedusing GeneSpring™ software analysis as described herein. Each columnrepresents the hybridization pattern resulting from a single individual.Normal individuals have no known medical conditions and were not takingany known medication. Non depression individuals presented withoutdepression, but may have presented with other medical conditions and maybe under various treatment regimes. Hybridizations to create said geneexpression profiles were done using ChondroChip™. A dendogram analysisis shown above. Samples are clustered and marked as representingpatients who have depression, having non-depression or normal. The “*”indicates those patients who abnormally clustered despite actualpresentation. The number of hybridizations profiles determined forpatients with depression, non-depression and normal are shown. 941 geneswere identified as being differentially expressed with a p value of<0.05 as between the patients with depression and a combination of thoseindividuals without depression and normal is noted. The identity of thedifferentially expressed genes is shown in Table 3N.

[0382] Classification or class prediction of a test sample of anindividual to determine whether said individuals has depression or doesnot having depression can be done using the differentially expressedgenes as shown in Table 3N as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 24

[0383] ChondroChip™ Microarray Data Analysis of gene expression profilesof blood samples from individuals having osteoarthritis as compared withgene expression profiles from normal individuals.

[0384] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswho were identified as having various stages of osteoarthritis ascompared to blood samples taken from healthy patients.

[0385] Osteoarthritis (OA), as used herein also known as “degenerativejoint disease”, represents failure of a diarthrodial (movable,synovial-lined) joint. It is a condition, which affects joint cartilage,and or subsequently underlying bone and supporting tissues leading topain, stiffness, movement problems and activity limitations. It mostoften affects the hip, knee, foot, and hand, but can affect other jointsas well.

[0386] OA severity can be graded according to the system described byMarshall (Marshall, K. W., J. Rheumatol., 1996, 23(4):582-85). Briefly,each of the six knee articular surfaces was assigned a cartilage gradewith points based on the worst lesion seen on each particular surface.Grade 0 is normal (0 points), Grade I cartilage is soft or swollen butthe articular surface is intact (1 point). In Grade II lesions, thecartilage surface is not intact but the lesion does not extend down tosubchondral bone (2 points). Grade III damage extends to subchondralbone but the bone is neither eroded nor eburnated (3 points). In GradeIV lesions, there is eburnation of or erosion into bone (4 points). Aglobal OA score is calculated by summing the points from all sixcartilage surfaces. If there is any associated pathology, such asmeniscus tear, an extra point will be added to the global score. Basedon the total score, each patient is then categorized into one of four OAgroups: mild (1-6), moderate (7-12), marked (13-18), and severe (>18).As used herein, patients identified with OA may be categorized in any ofthe four OA groupings as described above.

[0387] Blood samples were taken from patients who were diagnosed withosteoarthritis and a specific stage of osteoarthritis as defined herein.Gene expression profiles were then analysed and compared to profilesfrom patients unaffected by any disease. In each case, the diagnosis ofosteoarthritis and the stage of osteoarthritis was corroborated by askilled Board certified physician.

[0388] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a 15K Chondrogene MicroarrayChip (ChondroChip™) as described herein. Identification of genesdifferentially expressed in blood samples from patients with disease ascompared to healthy patients was determined by statistical analysisusing the Wilcox Mann Whitney rank sum test (Glantz S A., Primer ofBiostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0389]FIG. 22 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals having osteoarthritis ascompared with gene expression profiles from normal individuals.Expression profiles were generated using GeneSpring™ software analysisas described herein. Each column represents the hybridization patternresulting from a single individual. Normal individuals have no knownmedical conditions and were not taking any known medication.Hybridizations to create said gene expression profiles were done usingthe ChondroChip™. A dendogram analysis is shown above. Samples areclustered and marked as representing patients who presented withdifferent stages of osteoarthritis or normal. The “*” indicates thosepatients who abnormally clustered despite actual presentation. Thenumber of hybridizations profiles determined for either osteoarthritispatients or normal individuals are shown. 300 differentially expressedgenes were identified as being differentially expressed with a p valueof <0.05 as between the osteoarthritis patients and normal individuals.The identity of the differentially expressed genes is shown in Table 30.

[0390] Classification or class prediction of a test sample of anindividual as having OA, having mild OA, having marked OA, havingmoderate OA, having severe OA or not having OA can be done using thedifferentially expressed genes as shown in Table 30 as the predictorgenes in combination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 25

[0391] Microarray Data Analysis of gene expression profiles of bloodsamples from individuals having a condition as compared with geneexpression profiles from individuals not having said condition, andwherein said individual is undergoing therapeutic treatment in light ofsaid condition.

[0392] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken fromindividuals undergoing therapeutic treatment of a condition as comparedwith gene expression profiles from individuals not undergoing treatment.

[0393] Blood samples are taken from patients who are undergoingtherapeutic treatment. Gene expression profiles are then analysed andcompared to profiles from patients not undergoing treatment.

[0394] Total mRNA from a drop of peripheral whole blood taken from eachpatient is isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample are generated as described above.Each probe is denatured and hybridized to a microarray for example the15K Chondrogene Microarray Chip (ChondroChip™), Affymetrix Genechip orBlood chip as described herein. Identification of genes differentiallyexpressed in blood samples from patients undergoing therapeutictreatment as compared to patients not undergoing treatment is determinedby statistical analysis using the Wilcox Mann Whitney rank sum test(Glantz S A., Primer of Biostatistics. 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002). Expression profiles aregenerated using GeneSpring™ software analysis as described herein. Thenumber of differentially expressed genes are then identified as beingdifferentially expressed with a p value of <0.05.

EXAMPLE 26

[0395] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having liver cancer ascompared with gene expression profiles from normal individuals.

[0396] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith liver cancer as compared to blood samples taken from healthypatients.

[0397] As used herein, “liver cancer” means primary liver cancer whereinthe cancer initiates in the liver. Primary liver cancer includes bothhepatomas or hepatocellular carcinomas (HCC) which start in the liverand chonalgiomas where cancers develop in the bile ducts of the liver.

[0398] Blood samples were taken from patients who were diagnosed withliver cancer as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of liver cancer was corroborated bya skilled Board certified physician.

[0399] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with liver cancer as compared to healthypatients was determined by statistical analysis using the Weltch t-Test.

[0400]FIG. 25 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingliver cancer as described herein as compared with gene expressionprofiles from non-liver cancer disease individuals. Expression profileswere generated using GeneSpring™ software analysis as described herein.Each column represents the hybridization pattern resulting from a singleindividual. Control samples presented without liver cancer but may havepresented with other medical conditions and may be under varioustreatment regimes. Hybridizations to create said gene expressionprofiles were done using the Affymetrix™ U133A chip. A dendogramanalysis is shown above. Samples are clustered and marked asrepresenting patients who have liver cancer or control. The number ofhybridizations profiles determined for patients with liver cancer or whoare controls are shown. 1,475 genes were identified as beingdifferentially expressed with a p value of <0.05 as between the livercancer patients and those control individuals. The identity of thedifferentially expressed genes is shown in Table 3×.

[0401] Classification or class prediction of a test sample of anindividual to determine whether said individual has liver cancer or doesnot have liver cancer can be done using the differentially expressedgenes as shown in Table 3× as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 27

[0402] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having schizophrenia ascompared with gene expression profiles from normal individuals.

[0403] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith schizophrenia as compared to blood samples taken from healthypatients.

[0404] As used herein, “schizophrenia” is defined as a psychoticdisorders characterized by distortions of reality and disturbances ofthought and language and withdrawal from social contact. Patientsdiagnosed with “schizophrenia” can include patients having any of thefollowing diagnosis: an acute schizophrenic episode, borderlineschizophrenia, catatonia, catatonic schizophrenia, catatonic typeschizophrenia, disorganized schizophrenia, disorganized typeschizophrenia, hebephrenia, hebephrenic schizophrenia, latentschizophrenia, paranoic type schizophrenia, paranoid schizophrenia,paraphrenia, paraphrenic schizophrenia, psychosis, reactiveschizophrenia or the like.

[0405] Blood samples were taken from patients who were diagnosed withschizophrenia as defined herein. Gene expression profiles were thenanalysed and compared to profiles from patients unaffected by anydisease. In each case, the diagnosis of schizophrenia was corroboratedby a skilled Board certified physician.

[0406] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with schizophrenia as compared to healthypatients was determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A., Primer of Biostatistics, 5th ed.,New York, USA: McGraw-Hill Medical Publishing Division).

[0407]FIG. 26 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingschizophrenia as described herein as compared with gene expressionprofiles from non schizophrenic individuals. Expression profiles weregenerated using GeneSpring™ software analysis as described herein. Eachcolumn represents the hybridization pattern resulting from a singleindividual. Control samples presented without schizophrenia but may havepresented with other medical conditions and may be under varioustreatment regimes. Hybridizations to create said gene expressionprofiles were done using the Affymetrix™ U133A chip. A dendogramanalysis is shown above. Samples are clustered and marked asrepresenting patients who have schizophrenia or control individuals. Thenumber of hybridizations profiles determined for patients with livercancer or who are controls are shown. 1,952 genes were identified asbeing differentially expressed with a p value of <0.05 as between theschizophrenic patients and those control individuals. The identity ofthe differentially expressed genes is shown in Table 3Y.

[0408] Classification or class prediction of a test sample of anindividual to determine whether said individual has schizophrenia ordoes not having schizophrenia can be done using the differentiallyexpressed genes as shown in Table 3Y as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 28

[0409] Affymetrix U133A Chip Microarray Data Analysis of gene expressionprofiles of blood samples from individuals having Chagas disease ascompared with gene expression profiles from normal individuals.

[0410] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientswith symptomatic Chagas disease, asymptomatic Chagas disease or controlindividuals wherein said control individuals were confirmed as nothaving Chagas disease.

[0411] As used herein, “Chagas disease” is defined as a conditionwherein an individual is infected with the protozoan parasiteTrypanosoma cruzi and includes both acute and chronic infection. Acuteinfection with T cruzi can be diagnosed by detection of parasites byeither microscopic examination of fresh anticoagulated blood or thebuffy coat, giemsa-stained thin and thick blood smears and/or mouseinoculation and culturing of the blood of a potentially infectedindividual. Even in the absence of a positive result from the above, anaccurate determination of infection can be made by xenodiagnosis whereinreduviid bugs are allowed to feed on the patient's blood andsubsequently the bugs are examined for infection. Chronic infection canbe determined by detection of antibodies specific to the T cruziantigens and/or immunoprecipitation and electrophoresis of the T cruziantigens.

[0412] As used herein “Symptomatic Chagas disease” includes symptomaticacute chagas and symptomatic chronic chagas disease. Acute symptomaticchagas disease can be characterized by one or more of the following:area of erythema and swelling (a chagoma); local lymphadenopathy;generalized lymphadenopathy; mild hepatosplenomegaly; unilateralpainless edema of the palpebrae and periocular tissues; malaise; fever;anorexia and/or edema of the face and lower extremities. Symptomaticchronic Chagas' disease includes one or more of the following symptoms:heart rhythm disturbances, cardiomyopathy, thromboembolism,electrocardiographic abnormalities including right bundle-branchblockage; atrioventricular block; premature ventricular contractions andtachy- and bradyarrhythmias; dysphagia; odynophagia, chest pain;regurgitation; weight loss, cachexia and pulmonary infections.

[0413] As used herein “Asymptomatic Chagas disease” is meant to refer toindividuals who are infected with T cruzi but who do not show eitheracute or chronic symptoms of the disease.

[0414] Blood samples were taken from patients who were diagnosedsymptomatic or asymptomatic Chagas disease as defined herein. Geneexpression profiles were then analysed and compared to profiles frompatients unaffected by any disease. In each case, the diagnosis ofChagas disease was corroborated by a qualified physician.

[0415] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to an Affymetrix U133A Chip asdescribed herein. Identification of genes differentially expressed inblood samples from patients with Chagas disease as compared to healthypatients was determined by statistical analysis using the Weltch ANOVAtest (Michelson and Schofield, 1996).

[0416]FIG. 27 shows a diagrammatic representation of gene expressionprofiles of blood samples from individuals who were identified as havingsymptomatic Chagas disease; asymptomatic Chagas disease or who werecontrol individuals as described herein as compared with gene expressionprofiles from non-schizophrenic individuals. Expression profiles weregenerated using GeneSpring™ software analysis as described herein. Eachcolumn represents the hybridization pattern resulting from a singleindividual. Control samples presented without Chagas disease but mayhave presented with other medical conditions and may be under varioustreatment regimes.

[0417] Hybridizations to create said gene expression profiles were doneusing the Affymetrix™ U133A chip. A dendogram analysis is shown above.Samples are clustered and marked as representing patients who havesymptomatic chagas disease; asymptomatic chagas disease or control. Thenumber of hybridizations profiles determined for patients with chagasdisease; asymptomatic chagas disease or who are controls are shown. 668genes were identified as being differentially expressed with a p valueof <0.05 as between the symptomatic, asymptomatic Chagas patients andthose control individuals. The identity of the differentially expressedgenes is shown in Table 3Y.

[0418] Classification or class prediction of a test sample of anindividual to determine whether said individual has symptomatic Chagasdisease, asymptomatic Chagas disease or does not have Chagas disease canbe done using the differentially expressed genes as shown in Table 3Y asthe predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 29

[0419] Identification of Genes Specific for OA Only by Removing GenesRelevant to Co-Morbidities and Other Disease States.

[0420] This example demonstrates the use of the claimed invention todetect differential gene expression in blood unique to Osteoarthritis ascompared with other disease states.

[0421] Blood samples were taken from patients who were diagnosed withmild OA or severe OA and compared with individuals who were identifiedas normal individuals as defined herein. Gene expression profiles werethen analysed to identify genes which are differentially expressed in OAas compared with normal. In each case, the diagnosis of OA wascorroborated by a qualified physician.

[0422] Total mRNA from a drop of peripheral whole blood taken from eachpatient was isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample were generated as described above.Each probe was denatured and hybridized to a ChondroChip™ as describedherein. Identification of genes differentially expressed in bloodsamples from patients with mild or severe OA as compared to healthypatients was determined by statistical analysis using the Weltch ANOVAtest (Michelson and Schofield, 1996). (Dendogram analysis not shown).

[0423] In order to identify genes differentially expressed in bloodunique to OA but not differentially expressed as a result of possibleco-morbidities including hypertension, obesity, asthma, taking systemicsteroids, or allergies, genes identified as differentially expressed inboth OA and any of the genes identified as differentially expressed as aresult of co-morbidity, e.g., Table 3A (co-morbidity of OA andhypertension v. normal), Table 3B (co-morbidity of OA and obesity v.normal), Table 3C (co-morbidity of OA and allergy v. normal), Table 3D(co-morbidity of OA and taking systemic steroids v. normal), and genesin common with people identified as having asthma and OA (Table 3AA)were removed. Similarly any genes and unique to obesity (Table 3R),hypertension (Table 3P), allergies (Table 3T), systemic steroids (Table3V) were also removed. As a result of these comparisons, a list of genesunique to individuals with OA was identified. The identity of thedifferentially expressed genes is shown in Table 3AB.

[0424] It would be clear to a person skilled in the art that rather thansimply remove those genes which are relevant to other disease states,one could use a more refined analysis and remove those genes which showthe same trend in gene expression, e.g. remove those genes which show upregulation in a co-morbid state and also show up-regulation in thesingle disease state, but retain those genes which show a differenttrend in gene expression e.g. retain those genes which show upregulation in a co-morbid state as compared to down regulation in asingle disease state.

[0425] Classification or class prediction of a test sample of anindividual to determine whether said individual has OA or does not haveOA can be done using the differentially expressed genes as shown inTable 3AB, irrespective of whether the individual presents withco-morbidity using well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 30

[0426] Analysis of gene expression profiles of blood samples fromindividuals having brain cancer as compared with gene expressionprofiles from normal individuals.

[0427] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with brain cancer as compared to blood samples taken fromhealthy patients.

[0428] As used herein “brain cancer” refers to all forms of primarybrain tumours, both intracranial and extracranial and includes one ormore of the following: Glioblastoma, Ependymoma, Gliomas, Astrocytoma,Medulloblastoma, Neuroglioma, Oligodendroglioma, Meningioma,Retinoblastoma, and Craniopharyngioma.

[0429] Blood samples are taken from patients diagnosed with brain canceras defined herein. Gene expression profiles are then analysed andcompared to profiles from patients unaffected by any disease. Preferablyhealthy patients are chosen who are age and sex matched to said patientsdiagnosed with disease. In each case, the diagnosis of brain cancer iscorroborated by a skilled Board certified physician.

[0430] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample are generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients with braincancer as compared to healthy patients is determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A.,Primer of Biostatistics, 5th ed., New York, USA: McGraw-Hill MedicalPublishing Division, 2002).

[0431] Classification or class prediction of a test sample of anindividual to determine whether said individuals has brain cancer ordoes not having brain cancer can be done using the differentiallyexpressed genes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 31

[0432] Analysis of gene expression profiles of blood samples fromindividuals having ankylosing spondylitis as compared with geneexpression profiles from normal individuals.

[0433] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with ankylosing spondylitis as compared to blood samples takenfrom healthy patients.

[0434] As used herein “ankylosing spondylitis” refers to a chronicinflammatory disease that affects the joints between the vertebrae ofthe spine, and/or the joints between the spine and the pelvis and caneventually cause the affected vertebrae to fuse or grow together.

[0435] Blood samples are taken from patients diagnosed with ankylosingspondylitis as defined herein. Gene expression profiles are thenanalysed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of ankylosing spondylitis is corroborated by a skilled Boardcertified physician.

[0436] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withankylosing spondylitis as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A, Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0437] Classification or class prediction of a test sample of anindividual to determine whether said individuals has ankylosingspondylitis or does not having ankylosing spondylitis can be done usingthe differentially expressed genes identified as described above as thepredictor genes in combination with well known statistical algorithms aswould be understood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 32

[0438] Analysis of gene expression profiles of blood samples fromindividuals having prostate cancer as compared with gene expressionprofiles from normal individuals.

[0439] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with prostate cancer as compared to blood samples taken fromhealthy patients

[0440] As used herein “prostate cancer” refers to a malignant canceroriginating within the prostate gland. Patients identified as havingprostate cancer can have any stage of prostate cancer, as determinedclinically (by digital rectal exam or PSA testing) and orpathologically. Staging of prostate cancer can done in accordance withTNM or the Staging System of the American Joint Committee on Cancer(AJCC). In addition to the TNM system, other systems may be used tostage prostate cancer, for example, the Whitmore-Jewett system.

[0441] Blood samples are taken from patients diagnosed with prostatecancer as defined herein. Gene expression profiles are then analysed andcompared to profiles from patients unaffected by any disease to identifygenes which differentiate as between the two groups. Similarly geneexpression profiles can be analysed so as to differentiate as betweenthe severity of the prostate cancer. Preferably healthy patients arechosen who are age and sex matched to said patients diagnosed withdisease or with a specific stage of said disease. In each case, thediagnosis of prostate cancer is corroborated by a skilled Boardcertified physician.

[0442] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChipT as described herein. Identification ofgenes differentially expressed in blood samples from patients withprostate cancer as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A., Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0443] Classification or class prediction of a test sample of anindividual to determine whether said individuals has prostate cancer,has a specific stage of prostate cancer, or does not having prostatecancer can be done using the differentially expressed genes identifiedas described above as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics (e.g. GeneSpring™) for Class Predicationare also available.

EXAMPLE 33

[0444] Analysis of gene expression profiles of blood samples fromindividuals having ovarian cancer as compared with gene expressionprofiles from normal individuals.

[0445] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with ovarian cancer as compared to blood samples taken fromhealthy patients.

[0446] As used herein “ovarian cancer” refers to a malignant cancerousgrowth originating within the ovaries. Patients identified as havingovarian cancer can have any stage of ovarian cancer. Staging is done bycombining information from imaging tests with the results of a surgicalexamination done during a laprotomy. Numbered stages I to IV are used todescribe the extent of the cancer and whether it has spread(metastasized) to more distant organs.

[0447] Blood samples are taken from patients diagnosed with ovariancancer, or with a specific stage of ovarian cancer as defined herein.Gene expression profiles are then analysed and compared to profiles frompatients unaffected by any disease. Preferably healthy patients arechosen who are age and sex matched to said patients diagnosed withdisease or with a specific stage of said disease. In each case, thediagnosis of ovarian cancer is corroborated by a skilled Board certifiedphysician.

[0448] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withovarian cancer and or a specific stage of ovarian cancer as compared tohealthy patients is determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A., Primer of Biostatistics, 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0449] Classification or class prediction of a test sample of anindividual to determine whether said individuals has ovarian cancer, hasa specific stage of ovarian cancer or does not having ovarian cancer canbe done using the differentially expressed genes identified as describedabove as the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 34

[0450] Analysis of gene expression profiles of blood samples fromindividuals having kidney cancer as compared with gene expressionprofiles from normal individuals.

[0451] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with kidney cancer as compared to blood samples taken fromhealthy patients.

[0452] As used herein “kidney cancer” refers to a malignant cancerousgrowth originating within the kidneys. Kidney cancer includes renal cellcarcinoma, transitional cell carcinoma, and

[0453] Wilms' tumor. Patients identified as having renal cell carcinomacan also be categorized by stage of said cancer as determined by theSystem of the American Joint Committee on Cancer (AJCC). Numbered stagesI to IV are used to describe the extent of the carcinoma and whether ithas spread (metastased) to more distant organs.

[0454] Blood samples are taken from patients diagnosed with kidneycancer, or with a specific stage of renal cell carcinoma as definedherein. Gene expression profiles are then analysed and compared toprofiles from patients unaffected by any disease. Preferably healthypatients are chosen who are age and sex matched to said patientsdiagnosed with disease or with a specific stage of said disease. In eachcase, the diagnosis of kidney cancer is corroborated by a skilled Boardcertified physician.

[0455] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChipT as described herein. Identification ofgenes differentially expressed in blood samples from patients withkidney cancer and or a specific stage of kidney cancer as compared tohealthy patients is determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A, Primer of Biostatistics, 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0456] Classification or class prediction of a test sample of anindividual to determine whether said individuals has kidney cancer, hasa specific stage of kidney cancer or does not having kidney cancer canbe done using the differentially expressed genes identified as describedabove as the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 35

[0457] Analysis of gene expression profiles of blood samples fromindividuals having gastric cancer as compared with gene expressionprofiles from normal individuals.

[0458] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with gastric cancer as compared to blood samples taken fromhealthy patients.

[0459] As used herein “gastric or stomach cancer” refers to a cancerousgrowth originating within the stomach and includes gastricadenocarcinoma, primary gastric lymphoma and gastric nonlymphoidsarcoma. Patients identified as having stomac can also be categorized bystage of said cancer as determined by the System of the American JointCommittee on Cancer (AJCC).

[0460] Blood samples are taken from patients diagnosed with stomachcancer, or with a specific stage of stomach cancer as defined herein.Gene expression profiles are then analysed and compared to profiles frompatients unaffected by any disease. Preferably healthy patients arechosen who are age and sex matched to said patients diagnosed withdisease or with a specific stage of said disease. In each case, thediagnosis of stomach cancer is corroborated by a skilled Board certifiedphysician.

[0461] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChipT as described herein. Identification ofgenes differentially expressed in blood samples from patients withstomach cancer and or a specific stage of stomach cancer as compared tohealthy patients is determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A, Primer of Biostatistics, 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0462] Classification or class prediction of a test sample of anindividual to determine whether said individuals has stomach cancer, hasa specific stage of stomach cancer or does not having stomach cancer canbe done using the differentially expressed genes identified as describedabove as the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 36

[0463] Analysis of gene expression profiles of blood samples fromindividuals having lung cancer as compared with gene expression profilesfrom normal individuals.

[0464] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with lung cancer as compared to blood samples taken fromhealthy patients.

[0465] As used herein “lung cancer” refers to a cancerous growthoriginating within the lung and includes adenocarcinoma, alveolar cellcarcinoma, squamous cell carcinoma, large cell and small cellcarcinomas. Patients identified as having lung cancer can also becategorized by stage of said cancer as determined by the System of theAmerican Joint Committee on Cancer (AJCC).

[0466] Blood samples are taken from patients diagnosed with lung cancer,or with a specific stage of lung cancer as defined herein. Geneexpression profiles are then analysed and compared to profiles frompatients unaffected by any disease. Preferably healthy patients arechosen who are age and sex matched to said patients diagnosed withdisease or with a specific stage of said disease. In each case, thediagnosis of lung cancer is corroborated by a skilled Board certifiedphysician.

[0467] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients with lungcancer and or a specific stage of lung cancer as compared to healthypatients is determined by statistical analysis using the Wilcox MannWhitney rank sum test (Glantz S A, Primer of Biostatistics, 5th ed., NewYork, USA: McGraw-Hill Medical Publishing Division, 2002).

[0468] Classification or class prediction of a test sample of anindividual to determine whether said individuals has lung cancer, has aspecific stage of lung cancer or does not having lung cancer can be doneusing the differentially expressed genes identified as described aboveas the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 37

[0469] Analysis of gene expression profiles of blood samples fromindividuals having breast cancer as compared with gene expressionprofiles from normal individuals.

[0470] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with breast cancer as compared to blood samples taken fromhealthy patients.

[0471] As used herein “breast cancer” refers to a cancerous growthoriginating within the breast and includes invasive and non invasivebreast cancer such as ductal carcinoma in situ (DCIS), lobular carcinomain situ (LCIS), infiltrating ductal carcinoma, and infiltrating lobularcarcinoma. Patients identified as having breast cancer can also becategorized by stage of said cancer as determined by the System of theAmerican Joint Committee on Cancer (AJCC) or TNM classification.

[0472] Blood samples are taken from patients diagnosed with breastcancer, or with a specific stage of breast cancer as defined herein.Gene expression profiles are then analysed and compared to profiles frompatients unaffected by any disease. Preferably healthy patients arechosen who are age and sex matched to said patients diagnosed withdisease or with a specific stage of said disease. In each case, thediagnosis of breast cancer is corroborated by a skilled Board certifiedphysician.

[0473] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withbreast cancer and or a specific stage of breast cancer as compared tohealthy patients is determined by statistical analysis using the WilcoxMann Whitney rank sum test (Glantz S A, Primer of Biostatistics, 5thed., New York, USA: McGraw-Hill Medical Publishing Division, 2002).

[0474] Classification or class prediction of a test sample of anindividual to determine whether said individuals has breast cancer, hasa specific stage of breast cancer or does not have breast cancer can bedone using the differentially expressed genes identified as describedabove as the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 38

[0475] Analysis of gene expression profiles of blood samples fromindividuals having nasopharyngeal cancer as compared with geneexpression profiles from normal individuals.

[0476] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with nasopharyngeal cancer as compared to blood samples takenfrom healthy patients.

[0477] As used herein “nasopharyngeal cancer” refers to a cancerousgrowth arising from the epithelial cells that cover the surface and linethe nasopharynx. Patients identified as having nasopharyngeal cancer canalso be categorized by stage of said cancer as determined by the Systemof the American Joint Committee on Cancer (AJCC) or TNM classification.

[0478] Blood samples are taken from patients diagnosed withnasopharyngeal cancer, or with a specific stage of nasopharyngeal canceras defined herein. Gene expression profiles are then analysed andcompared to profiles from patients unaffected by any disease. Preferablyhealthy patients are chosen who are age and sex matched to said patientsdiagnosed with disease or with a specific stage of said disease. In eachcase, the diagnosis of nasopharyngeal cancer is corroborated by askilled Board certified physician.

[0479] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to a AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withnasopharyngeal cancer and or a specific stage of breast cancer ascompared to healthy patients is determined by statistical analysis usingthe Wilcox Mann Whitney rank sum test (Glantz S A, Primer ofBiostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0480] Classification or class prediction of a test sample of anindividual to determine whether said individuals has nasopharyngealcancer, has a specific stage of nasopharyngeal cancer or does not havenasopharyngeal cancer can be done using the differentially expressedgenes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 39

[0481] Analysis of gene expression profiles of blood samples fromindividuals having Guillain Barre syndrome as compared with geneexpression profiles from normal individuals.

[0482] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Guillain Barre syndrome as compared to blood samplestaken from healthy patients.

[0483] As used herein “Guillain Barre syndrome” refers to an acute,usually rapidly progressive form of inflammatory polyneuropathycharacterized by muscular weakness and mild distal sensory loss.

[0484] Blood samples are taken from patients diagnosed with GuillainBarre syndrome as defined herein. Gene expression profiles are thenanalysed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of Guillain Barre syndrome is corroborated by a skilled Boardcertified physician.

[0485] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withGuillain Barre syndrome as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A, Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0486] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Guillain Barresyndrome, or does not have Guillain Barre syndrome can be done using thedifferentially expressed genes identified as described above as thepredictor genes in combination with well known statistical algorithms aswould be understood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 40

[0487] Analysis of gene expression profiles of blood samples fromindividuals having Fibromyalgia as compared with gene expressionprofiles from normal individuals.

[0488] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Fibromyalgia as compared to blood samples taken fromhealthy patients.

[0489] As used herein “Fibromyalgia” refers to widespread chronicmusculoskeletal pain and fatigue. The pain comes from the connectivetissues, such as the muscles, tendons, and ligaments and does notinvolve the joints. Blood samples are taken from patients diagnosed withFibromyalgia as defined herein. Gene expression profiles are thenanalysed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of Fibromyalgia is corroborated by a skilled Board certifiedphysician.

[0490] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withFibromyalgia as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A., Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0491] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Fibromyalgia, ordoes not have Fibromyalgia can be done using the differentiallyexpressed genes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 41

[0492] Analysis of gene expression profiles of blood samples fromindividuals having Multiple Sclerosis as compared with gene expressionprofiles from normal individuals.

[0493] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Multiple Sclerosis as compared to blood samples takenfrom healthy patients.

[0494] As used herein “Multiple Sclerosis” refers to chronic progressivenervous disorder involving the loss of myelin sheath surrounding certainnerve fibres. Blood samples are taken from patients diagnosed withMultiple Sclerosis as defined herein. Gene expression profiles are thenanalysed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of Multiple Sclerosis is corroborated by a skilled Boardcertified physician.

[0495] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withMultiple Sclerosis as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A, Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0496] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Multiple Sclerosis,or does not have Multiple Sclerosis can be done using the differentiallyexpressed genes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 42

[0497] Analysis of gene expression profiles of blood samples fromindividuals having Muscular Dystrophy as compared with gene expressionprofiles from normal individuals.

[0498] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Muscular Dystrophy as compared to blood samples takenfrom healthy patients.

[0499] As used herein “Muscular Dystrophy” refers to a hereditarydisease of the muscular system characterized by weakness and wasting ofthe skeletal muscles. Muscular Dystrophy includes Duchennes' MuscularDystrophy, limb-girdle muscular dystrophy, myotonia atrophica, myotonicmuscular dystrophy, pseudohypertrophic muscular dystrophy, andSteinhardt's disease.

[0500] Blood samples are taken from patients diagnosed with MuscularDystrophy as defined herein. Gene expression profiles are then analysedand compared to profiles from patients unaffected by any disease.Preferably healthy patients are chosen who are age and sex matched tosaid patients diagnosed with disease. In each case, the diagnosis ofMuscular Dystrophy is corroborated by a skilled Board certifiedphysician.

[0501] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withMuscular Dystrophy as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A, Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0502] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Muscular Dystrophy,or does not have Muscular Dystrophy can be done using the differentiallyexpressed genes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 43

[0503] Analysis of gene expression profiles of blood samples fromindividuals having septic joint arthroplasty as compared with geneexpression profiles from normal individuals.

[0504] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with septic joint arthroplasty as compared to blood samplestaken from healthy patients.

[0505] As used herein “septic joint arthroplasty” refers to aninflammation of the joint caused by a bacterial infection.

[0506] Blood samples are taken from patients diagnosed with septic jointarthroplasty as defined herein. Gene expression profiles are thenanalysed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of septic joint arthroplasty is corroborated by a skilledBoard certified physician.

[0507] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withseptic joint arthroplasty as compared to healthy patients is determinedby statistical analysis using the Wilcox Mann Whitney rank sum test(Glantz S A, Primer of Biostatistics, 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0508] Classification or class prediction of a test sample of anindividual to determine whether said individuals has septic jointarthroplasty, or does not have septic joint arthroplasty can be doneusing the differentially expressed genes identified as described aboveas the predictor genes in combination with well known statisticalalgorithms as would be understood by a person skilled in the art anddescribed herein. Commercially available programs such as those providedby Silicon Genetics (e.g. GeneSpring™) for Class Predication are alsoavailable.

EXAMPLE 44

[0509] Analysis of gene expression profiles of blood samples fromindividuals having Alzheimers Disease as compared with gene expressionprofiles from normal individuals.

[0510] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Alzheimers as compared to blood samples taken fromhealthy patients.

[0511] As used herein “Alzheimers” refers to a degenerative disease ofthe central nervous system characterized especially by premature senilemental deterioration.

[0512] Blood samples are taken from patients diagnosed with Alzheimersas defined herein. Gene expression profiles are then analysed andcompared to profiles from patients unaffected by any disease. Preferablyhealthy patients are chosen who are age and sex matched to said patientsdiagnosed with disease. In each case, the diagnosis of Alzheimers iscorroborated by a skilled Board certified physician.

[0513] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withAlzheimers as compared to healthy patients is determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A, Primerof Biostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0514] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Alzheimers, or doesnot have Alzheimers can be done using the differentially expressed genesidentified as described above as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 45

[0515] Analysis of gene expression profiles of blood samples fromindividuals having hepatitis as compared with gene expression profilesfrom normal individuals.

[0516] This example demonstrates the use of the claimed invention todetect gene expression in blood samples taken from patients diagnosedwith hepatitis as compared to blood samples taken from healthy patients.

[0517] As used herein “hepatitis” refers to an inflammation of the livercaused by a virus or toxin and can include hepatitis A, hepatitis B,hepatitis C, hepatitis D, hepatitis E, and hepatitis F.

[0518] Blood samples are taken from patients diagnosed with hepatitis asdefined herein. Gene expression profiles are then analysed and comparedto profiles from patients unaffected by any disease. Preferably healthypatients are chosen who are age and sex matched to said patientsdiagnosed with disease. In each case, the diagnosis of hepatitis iscorroborated by a skilled Board certified physician.

[0519] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU 133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withhepatitis as compared to healthy patients is determined by statisticalanalysis using the Wilcox Mann Whitney rank sum test (Glantz S A, Primerof Biostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0520] Classification or class prediction of a test sample of anindividual to determine whether said individuals has hepatitis, or doesnot have hepatitis can be done using the differentially expressed genesidentified as described above as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 46

[0521] Analysis of gene expression profiles of blood samples fromindividuals having Manic Depression Syndrome (MDS) as compared with geneexpression profiles from normal individuals.

[0522] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with MDS as compared to blood samples taken from healthypatients.

[0523] As used herein “Manic Depression Syndrome (MDS)” refers to a mooddisorder characterized by alternating mania and depression.

[0524] Blood samples are taken from patients diagnosed with MDS asdefined herein. Gene expression profiles are then analysed and comparedto profiles from patients unaffected by any disease. Preferably healthypatients are chosen who are age and sex matched to said patientsdiagnosed with disease. In each case, the diagnosis of MDS iscorroborated by a skilled Board certified physician.

[0525] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients with MDSas compared to healthy patients is determined by statistical analysisusing the Wilcox Mann Whitney rank sum test (Glantz S A, Primer ofBiostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0526] Classification or class prediction of a test sample of anindividual to determine whether said individuals has MDS, or does nothave MDS can be done using the differentially expressed genes identifiedas described above as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics (e.g. GeneSpring™) for Class Predicationare also available.

EXAMPLE 47

[0527] Analysis of gene expression profiles of blood samples fromindividuals having Crohn's Disease and/or Colitis as compared with geneexpression profiles from normal individuals.

[0528] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Crohn's Disease and/or Colitis as compared to bloodsamples taken from healthy patients.

[0529] As used herein “Crohn's Disease” refers to a chronic inflammationof the ileum which is often progressive. As used herein “Colitis” or“Inflammatory Bowel Disease” refers to inflammation of the colon.

[0530] Blood samples are taken from patients diagnosed with Crohn's andor Colitis as defined herein. Gene expression profiles are then analysedand compared to profiles from patients unaffected by any disease.Preferably healthy patients are chosen who are age and sex matched tosaid patients diagnosed with disease. In each case, the diagnosis ofCrohn's and or Colitis is corroborated by a skilled Board certifiedphysician.

[0531] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withCrohn's and or Colitis as compared to healthy patients is determined bystatistical analysis using the Wilcox Mann Whitney rank sum test (GlantzS A, Primer of Biostatistics, 5th ed., New York, USA: McGraw-HillMedical Publishing Division, 2002).

[0532] Classification or class prediction of a test sample of anindividual to determine whether said individuals has Crohn's and orColitis, or does not have Crohn's and or Colitis can be done using thedifferentially expressed genes identified as described above as thepredictor genes in combination with well known statistical algorithms aswould be understood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

EXAMPLE 48

[0533] Analysis of gene expression profiles of blood samples fromindividuals having Malignant Hyperthermia Susceptibility as comparedwith gene expression profiles from normal individuals.

[0534] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with Malignant Hyperthermia Susceptibility as compared toblood samples taken from healthy patients.

[0535] As used herein “Malignant Hyperthermia Susceptibility” refers toa pharmacogenetic disorder of skeletal muscle calcium regulation oftendeveloping during or after a general anaesthesia.

[0536] Blood samples are taken from patients diagnosed with MalignantHyperthermia Susceptibility as defined herein. Gene expression profilesare then analysed and compared to profiles from patients unaffected byany disease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of Malignant Hyperthermia Susceptibility is corroborated by askilled Board certified physician.

[0537] Total mRNA from a drop of peripheral whole blood is taken fromeach patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients withMalignant Hyperthermia Susceptibility as compared to healthy patients isdetermined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A, Primer of Biostatistics, 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002).

[0538] Classification or class prediction of a test sample of anindividual to determine whether said individuals has MalignantHyperthermia Susceptibility, or does not have Malignant HyperthermiaSusceptibility can be done using the differentially expressed genesidentified as described above as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 49

[0539] Analysis of gene expression profiles of blood samples from horseshaving osteoarthritis as compared with gene expression profiles fromnormal or non-osteoarthritic horses.

[0540] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from horsesso as to diagnose equine arthritis as compared to blood samples takenfrom healthy horses.

[0541] As used herein “arthritis” in reference to horses refers to adegenerative joint disease that affects horses by causing lameness.Although it can appear in any joint, most common areas are the upperknee joint, front fetlocks, hocks, or coffin joints in the front feet.The condition can be caused by trauma, mineral or dietary deficiency,old age, poor conformation, over exertion or infection. The differentstructures that can be damaged in arthritis are the cartilage insidejoints, the bone in the joints, the joint capsule, the synovialmembranes, the ligaments around the joints and lastly the fluid thatlubricates the insides of ‘synovial joints’. In severe cases all ofthese structures are affected. In for example osteochondrosis only thecartilage may be affected.

[0542] Regardless of the cause, the disease begins when the synovialfluid that lubricates healthy joints begins to thin. The decrease inlubrication causes the cartilage cushion to break down, and eventuallythe bones begin to grind painfully against each other. Diagnostic testsused to confirm arthritis include X-rays, joint fluid analysis, andultrasound.

[0543] Blood samples are taken from horses diagnosed with arthritis asdefined herein. Gene expression profiles are then analysed and comparedto profiles from horses unaffected by any disease. Preferably healthyhorses are chosen who are age and sex matched to said horses diagnosedwith disease. In each case, the diagnosis of arthritis is corroboratedby a certified veterinarian.

[0544] Total mRNA from a drop of peripheral whole blood is taken fromeach horse and isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample is generated as described above.Each probe is denatured and hybridized to an Affymetrix U133A Chipand/or a ChondroChip™ as described herein. An equine specific microarrayrepresenting the equine genome can also be used. Identification of genesdifferentially expressed in blood samples from horses with arthritis ascompared to healthy horses is determined by statistical analysis usingthe Wilcox Mann Whitney rank sum test (Glantz S A, Primer ofBiostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0545] Classification or class prediction of a test sample of a horse todetermine whether said horse has arthritis or does not have arthritiscan be done using the differentially expressed genes identified asdescribed above as the predictor genes in combination with well knownstatistical algorithms as would be understood by a person skilled in theart and described herein. Commercially available programs such as thoseprovided by Silicon Genetics (e.g. GeneSpring™) for Class Predicationare also available.

EXAMPLE 50

[0546] Analysis of gene expression profiles of blood samples from dogshaving osteoarthritis as compared with gene expression profiles fromnormal or non-osteoarthritic dogs.

[0547] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from dogs soas to diagnose equine arthritis as compared to blood samples taken fromhealthy horses.

[0548] As used herein “osteoarthritis” in reference to dogs is a form ofdegenerative joint disease which involves the deterioration of andchanges to the cartilage and bone. In response to inflammation in andabout the joint, the body responds with bony remodelling around thejoint structure. This process can be slow and gradual with minimaloutward symptoms, or more rapidly progressive with significant pain anddiscomfort. Osteoarthritic changes can occur in response to infectionand injury of the joint as well.

[0549] Blood samples are taken from dogs diagnosed with osteoarthritisas defined herein. Gene expression profiles are then analysed andcompared to profiles from dogs unaffected by any disease. Preferablyhealthy dogs are chosen who are age, sex and breed matched to said dogsdiagnosed with disease. In each case, the diagnosis of osteoarthritis iscorroborated by a certified veterinarian.

[0550] Total mRNA from a drop of peripheral whole blood is taken fromeach dog and isolated using TRIzol® reagent (GIBCO) and fluorescentlylabelled probes for each blood sample is generated as described above.Each probe is denatured and hybridized to an Affymetrix U133A Chipand/or a ChondroChip™ as described herein. A canine specific microarrayrepresenting the canine genome can also be used. Identification of genesdifferentially expressed in blood samples from dogs with osteoarthritisas compared to healthy horses is determined by statistical analysisusing the Wilcox Mann Whitney rank sum test (Glantz S A, Primer ofBiostatistics, 5th ed., New York, USA: McGraw-Hill Medical PublishingDivision, 2002).

[0551] Classification or class prediction of a test sample of a dog todetermine whether said dog has osteoarthritis or does not haveosteoarthritis can be done using the differentially expressed genesidentified as described above as the predictor genes in combination withwell known statistical algorithms as would be understood by a personskilled in the art and described herein. Commercially available programssuch as those provided by Silicon Genetics (e.g. GeneSpring™) for ClassPredication are also available.

EXAMPLE 51

[0552] Analysis of gene expression profiles of blood samples fromindividuals having Manic Depression Syndrome (MDS) as compared with geneexpression profiles from individuals having Schizophrenia.

[0553] This example demonstrates the use of the claimed invention todetect differential gene expression in blood samples taken from patientsdiagnosed with MDS as compared to blood samples taken from schizophrenicpatients.

[0554] As used herein “Manic Depression Syndrome (MDS)” refers to a mooddisorder characterized by alternating mania and depression. As usedherein, “schizophrenia” is defined as a psychotic disorderscharacterized by distortions of reality and disturbances of thought andlanguage and withdrawal from social contact. Patients diagnosed with“schizophrenia” can include patients having any of the followingdiagnosis: an acute schizophrenic episode, borderline schizophrenia,catatonia, catatonic schizophrenia, catatonic type schizophrenia,disorganized schizophrenia, disorganized type schizophrenia,hebephrenia, hebephrenic schizophrenia, latent schizophrenia, paranoictype schizophrenia, paranoid schizophrenia, paraphrenia, paraphrenicschizophrenia, psychosis, reactive schizophrenia or the like.

[0555] Blood samples are taken from patients diagnosed with MDS orSchizophrenia as defined herein. Gene expression profiles are thenanalyzed and compared to profiles from patients unaffected by anydisease. Preferably healthy patients are chosen who are age and sexmatched to said patients diagnosed with disease. In each case, thediagnosis of MDS and Schizophrenia is corroborated by a skilled Boardcertified physician. Total mRNA from a drop of peripheral whole blood istaken from each patient and isolated using TRIzol® reagent (GIBCO) andfluorescently labelled probes for each blood sample is generated asdescribed above. Each probe is denatured and hybridized to an AffymetrixU133A Chip and/or a ChondroChip™ as described herein. Identification ofgenes differentially expressed in blood samples from patients with MDSas compared to Schizophrenic patients as compared to normal individualsis determined by statistical analysis using the Wilcox Mann Whitney ranksum test (Glantz S A, Primer of Biostatistics, 5th ed., New York, USA:McGraw-Hill Medical Publishing Division, 2002) (data not shown). 294genes were identified as being differentially expressed with a p valueof <0.05 as between the schizophrenic patients, the MDS patients andthose control individuals. The identity of the differentially expressedgenes is shown in Table 3AC.

[0556] Classification or class prediction of a test sample of anindividual to determine whether said individuals has MDS, hasSchizophrenia or is normal can be done using the differentiallyexpressed genes identified as described above as the predictor genes incombination with well known statistical algorithms as would beunderstood by a person skilled in the art and described herein.Commercially available programs such as those provided by SiliconGenetics (e.g. GeneSpring™) for Class Predication are also available.

[0557] One skilled in the art will appreciate readily that the presentinvention is well adapted to carry out the objects and obtain the endsand advantages mentioned, as well as those objects, ends and advantagesinherent herein. The present examples, along with the methods,procedures, treatments, molecules, and specific compounds describedherein are presently representative of preferred embodiments, areexemplary, and are not intended as limitations on the scope of theinvention. Changes therein and other uses will occur to those skilled inthe art which are encompassed within the spirit of the invention asdefined by the scope of the claims.

[0558] All patents, patent applications, and published references citedherein are hereby incorporated by reference in their entirety. Whilethis invention has been particularly shown and described with referencesto preferred embodiments thereof, it will be understood by those skilledin the art that various changes in form and details may be made thereinwithout departing from the scope of the invention encompassed by theappended claims.

1 112 1 110 DNA Human 1 acacaacgta acaataacat atttagccaa tgtagtagactgctatataa tacattagag 60 tgtcaattca ttccgtttac agccccattg ggtgtcaaattttttttgtt 110 2 530 DNA Human misc_feature (16)..(16) n is a, c, g, ort 2 gcctgttcta tacagnttnt aaatntcatt tcagatcntn tntntgtgat aatgaatgct 60gttnnntagn natcctatat natgtncgna cacatcctaa agcataggat gaaaaantga 120nanccttagg atttngagca cantgccttt acctgaatat atacagcaca gttctgnant 180ncctggcgtg tgnnactgga gatctctann aaaangnata nagtgggngg gcnctntggc 240gcntgccggt nnnncctaaa ttttccccan gngnnggagg ccngtcacct gnncccatng 300cgntctngac cngcctgtna acgnntanng gagccttagt cnctnctaaa aacacaaaat 360tagccnggca tgggggntgg gncccttgta ntctnagctn cttgggaggc tnngccagga 420antncncttg aanccgggna gngggtggcc tnaagtttgn ggnaaggcca ntgatcaccg 480ccccttcccc tccangcccn gggngaaggg atttgngact tccgttttgg 530 3 215 DNAHuman 3 cggcacgagg atcaatttgc cttggaagaa caaaaggaaa gtctggaaatgcagaaagta 60 tggatgctga accacataac agcagatggc attgctgtga agtatactggatggaataca 120 ttcaagcgtt aatatttaat tctttttgtg gaaggtcaca caattaaaatttaattgggc 180 atggaggctt aggacggggt aaaaaagtct ttaga 215 4 129 DNAHuman 4 gtttcttttt cctaaaacgg ttttatttaa ctcaatgtgt caaagtttttttttaataat 60 cccaagaggg atgaagccgt gtccacaggg atatatacat cattatggttcccatctttc 120 atacatgaa 129 5 361 DNA Human misc_feature (13)..(14) nis a, c, g, or t 5 gggggctttt ttnnancggn nccgnnnncc cttcctgggaanttttgggc cnttntntna 60 aangnggnct tncnggnaaa tgggtttttt nagggggctggncaaaggtt ttttctntaa 120 tgggatnngg ccggcatttt aaaaaaaccc gctttggcctttttgctana tnggaaaaaa 180 tttttttaaa angcctaaga canggttttc ccttcatatgccaaactttc cctaacattt 240 ggnntttnng ggngggcagg gggggatttt taaaccggatttngggtnaa aaaaaatcng 300 gggggaattt ttgggganaa aaccttnggg gggnccccctttgaaaanaa agggtgggnn 360 g 361 6 839 DNA Human misc_feature(475)..(475) n is a, c, g, or t 6 ctcgtgccga attcggcacg agcaaagtacctggacttta tggaatcctt ctatacttca 60 ttgtcaatca tttattggtt ctaaaaaggatcggacaatg tgctatttca gggaagccaa 120 tgttttggag taaaatgcac aaataatttctcttgccttg caaacacatt tttttttctg 180 tcattgcaat gtgcacaaag ggccacgaggatctacaaga aagcctgcct tattctgacc 240 aggagtgggg agctgacaag aggcttcacagagcaggtga tgtttagaga ggaatgtctc 300 ccatttccta gtagcctgtg aggctctcaaaaccgggaat caagtttccc ttctgaactc 360 agttctcaat cgtgtaggga tagggttcccaggtgtgcct ctatgtgtag aggctctatt 420 ataccctgga tacacattga tatgcatgtgcaatgctgga atcaccagcc cccangtcct 480 cctcccaaat gtgcatgttt tttgacccatgtcacattta attttttttt tcaattgacg 540 ggtttttagg gcaaanttnc caaaacatcccccactttgc catantcccc tgtcattcca 600 tattgncttg cactgacatg attcactcattgatattgcc tgtngcgttc ctatggcctt 660 tgagtttgca nactgggttt gggggaaacccangnaaaaa aacctctttg aaanggggaa 720 cccccccaat ggtgggggaa ananaactggactttntttg ggagnccnga atttgctctt 780 gaccaggcag ggacctggga ccctgaangcttttntaatc ttnggggccn gaaaatntg 839 7 118 DNA Human 7 atgggaaagtgtgtaagatt tagaaaaagc attaactatt agtaaacttt atcttaagct 60 ctaacctttgattaggtccc acaaaaatta ggtgatatgc aatttctaat ttagggcc 118 8 197 DNA Human8 gttgcagtga gccgagatca taccactgca ctccagccta ggcaacagag cgagactcgg 60tcaaaagaaa aaaaaaaagg ggagctgggc gtgggtacta atgccgtaat cccaggcctt 120tgggaatccc aggcaaggtg gcctttaggg caaggagttc ggaacctccc tgctaacagg 180taaaccccct ttccctt 197 9 250 DNA Human 9 gagaccaagg ccgccccgctctggtctcag accagttgtg ctgctcttgc tctggctcag 60 ctggtgtggg gcgcaggcgggaaacgagac ctctagcatc tggctgaagg ctctgccaag 120 ctcctcttca gggctgcagtctgcctgcct gcatataccg acttggccag acactgctgc 180 taaattccag ggactctttctcccctcctc tgctctccag ccaatccttg aggatttaat 240 aactggaagg 250 10 680DNA Human misc_feature (433)..(433) n is a, c, g, or t 10 caccaaagaagcaagagggc tttcttttgt ttctggggac aataactaac tttaatttgc 60 tcttcaagaagaaggaagct gggtatatag gggaatggca gaagtgctcg cagatgaacc 120 atgaggagcatggtctttaa gaacatgctg agaaggaagc aacacagact ccatcactgg 180 gggaagcacctgaatagagc actggtaaag gccagtctgt ggacctgagg ccagaggaga 240 tgccaggggtccagatttca tggcccacag aaacggaact gatcatattt ggttgctggc 300 cagtgttccatagaccaaga aggctggtag caagtataga ttcctctaca tagcttgaca 360 ggagaagagaaaggggaatg tagcacacag gatgcagcag gtgaataaga aaacctcctt 420 ttcccaggttggngacagtg agtgatctac agtgatactc aaaagattgt gattggtgtg 480 ggaattcctgtctcaatatg caatctgcca agaaaacact gtgatggttt cctgtaaagt 540 aaccctcttttcttatctct aatttcacaa gactcttaaa tgagaggggg gggagaaagn 600 gttctttctcactcncctaa aactgngggt ctgcctggag aaaanctaca tctgcacaga 660 naatgctggttagccaggaa 680 11 318 DNA Human 11 cctgcagagt actccatgga aacaattgccgagcacgtgc tcgcaatttg ccgagcacgg 60 tccggtttga actcctagac taagactaggtaggtgatac ataccttctt cccaccaagt 120 actcacgatc caaactatga attttagattcggatcaaac gaggattgat ccgagggacc 180 aacgttgtga taaatcttac gtcgtcttatatattaagtt tttgtggagg atcggataag 240 tctatagtgt ttgtcacaga tagtcccgtaccacacccca gaccatagga gtcgctctcc 300 ggaccgcggt ctaatggg 318 12 155 DNAHuman 12 tctcacattg gacatactca aaattcactt ataatcttca caccaccaaaaacttaccca 60 tatcaaatta taaacccacc cacattactt aaaatttttt acatttcccaataaaaaacc 120 caaataaaca aaaacttcca atctccattt aaaat 155 13 125 DNAHuman 13 aataaacaaa catgccctct aatatatgaa ttcatcacac aacacgcacactgtccccac 60 aaacaccttt ttggtgtcaa gaagaaaaag actagcttca ctgaacagagaaatgctgga 120 cagtg 125 14 168 DNA Human misc_feature (6)..(6) n is a,c, g, or t 14 ggcccntggg ggggnagggc cttttcgggg ccggggnngg gcccccntttggcccnnggg 60 gggtttcccg gggaacccaa ccctttaagg ggtngggggg aatttccccccaaaaaaagg 120 gaaaaanttt tccggggggc ccacccggga agggntnccg gggaaggg 16815 438 DNA Human 15 aaaaaacttc tttatagtcc ttatatattt ttaattgtttatgttagggg aagctataga 60 ggaacaaatt tgggatagaa atataaggct gggattacaggcatgagcca ccaagcccgg 120 cccacatttc catttttaat atatactgtg ctttacaaatattataatat gttttaaaat 180 atgttcacag aagcacctgg tctgtgaatg gcatgccagcattaaaaaaa ataagcattc 240 tttgaatata tatttagttt tttaatgtgg taggaaaatcaaagccagag ggagtagaaa 300 caaaatttgt gattttctaa atacttcttg gctgcagggaagaaaccacg tcccaggcga 360 agtcctacct aatttgatga taaaattaca tggaagggattcttgttggc atgaggacct 420 accaagatgg tcaacaga 438 16 235 DNA Humanmisc_feature (5)..(5) n is a, c, g, or t 16 aaggnctttt ccggnccggcccggcccccc ttggcccang ggggttnccg gnaaaccacc 60 ctttaaggnt tgggggaattcccccaaaaa aggaaaaaat tttcccgggg gcccacccgg 120 aaagggggaa ggcccccaaaaccggggggg gggnaaaaag gtgggtttcc ccctttttcc 180 aattcccaaa accaatttccaaaaggnaaa ccaaccnttc ccaaaatggg aaagg 235 17 294 DNA Human misc_feature(18)..(19) n is a, c, g, or t 17 aaaccaaccc tttaaggnnt ggggggnaattccccccaaa aaaaggnaaa aattttttcc 60 gggggnccaa accggnaaag gntttgggaaaaccaaattt tttttggncc caaccccccc 120 caaattgggg ggnaaaccaa atttaaggggggaagggggg gncccccccg ggaaaggccc 180 aaggggggaa aatttttccg ggggtgggtngggggaacca atttaagggg ggggcccccg 240 ggggggttcc ccttgggccn tttttccttttgggtnaaaa aaaaaaaccc cttg 294 18 453 DNA Human 18 gtagaatata gggtgatactggagatctac tgcgacctag accatgatac ataaccacac 60 aagtttaatc cctgggttctaactaccctt actgtcactt agcttaacct gcctccaatc 120 ctgtacttga actctaaaactgttggagaa actcagtgct taccccaaca gattcatttc 180 aaatagctgt aaaaggtatgtttactccag aagaccagag ttgcttcttt tgaacttctc 240 attccttggg cctaggaaccctcatcaccc tcatcccaac gtcaacccag atcttctctt 300 ccataaacag cactccctcaggcccctgcc tgacacaggc atagactgtc atgttggatt 360 cacagacagg ctgtgctagaggaaacctct ggggctcacc aggggccgtg ggatgggctt 420 ctggggcttc ttggagcccaacttcttcat ggc 453 19 242 DNA Human misc_feature (17)..(17) n is a, c,g, or t 19 gagtcagact gtaaggnacg aaccctcggg gtccccacgn tgttccccccggggtaacnt 60 cggcccgggc ccgggnagcc cttcccgggc ttttcccccg ggggggncccgggggggacc 120 tttaggcggc accccaacaa caccaggccc tactttttcc aaggncggggaagcccatgg 180 gttctgggna acgggcaatg cgggcttgca acgggnggaa naaaaacagncccaaaagaa 240 tg 242 20 181 DNA Human 20 gtttgtttgt ttttgagatgaatctcactc tgtcgcccag gctggaatgc agtggtgtga 60 tctcagctca ctgcaacctccacctctcag gagaattgct gaacctggga ggcggaggtt 120 gcagggagct gagattgcgccactgccctc catcctgggc gacagagcaa gaacctgtct 180 c 181 21 100 DNA Humanmisc_feature (17)..(17) n is a, c, g, or t 21 gcacaaggaa gggtggncagatnttccngc actggnaaaa ngcngctatg gtngtgaant 60 tnccccnccn nttnanacnaaanntngcac tcttggntgc 100 22 100 DNA Human misc_feature (2)..(2) n is a,c, g, or t 22 cntgcgccat ttactgnagg tggacaagga tactatnaac aaagatgtggcnnaangaga 60 ataatggaag atagctntga ggatnaacnc tggttnaggg 100 23 100 DNAHuman misc_feature (17)..(17) n is a, c, g, or t 23 acaccttcccacttgcngna aaggggnnng gcccccnnct tgggcnganc attaagcctt 60 tttgnggctgcngcccctgt gcctggtgcc acaacaaatg 100 24 227 DNA Human misc_feature(5)..(5) n is a, c, g, or t 24 ccggncacca ccnttaaggt tgggggatttccccaaaaaa ggaaaatttt cggcggccaa 60 cgggaaggcc nttggggaaa aaaccaanggncaaaccccc ccaaccacnc ggcccccccc 120 aaggggggtg gggaagagcc aaatttctttgggaaanaac gcccccttgg ggaaaanaag 180 gccaaccacc tttcaacanc ccccaangcgnggaagccat ttcttgg 227 25 306 DNA Human 25 tccaaaagta gagcagagggatattttgtt ctactgagcc acgaaaaaca cctgaattgt 60 ttcgaccatg tgccttcccaggttgatgaa gacattgcta cacagtctgc agatcaggaa 120 ggaagaattg tatgtgggagtttttaatgg tctcatttca ttggctataa ctcagttaca 180 aggagaaata taactgcagaggagctttga aaatttagtt cagctgaggg taaaggaaga 240 agagacaaat tttgtcatcagctagtgatc tgccatacaa ggtgttccct taatatgtgt 300 agaatg 306 26 492 DNAHuman misc_feature (299)..(299) n is a, c, g, or t 26 cggcttcgggccaagcgttt ccagagtttg ccgaactgct gagcaagttc gctattctcc 60 agatcgcctagccctttgcg ggcgaccacc acgatgtccc agcctgtcag gttgtcctga 120 ttgaggcgaaaggactcgcg gatttgacgc ttgatgcggt tgcgctcgac ggcgagcttg 180 acgctctttttgccgatcac caaacctagg cggggatgat caagctggtt atcgcgcgct 240 agcagcaggacacttttgcc cgggagcttt accgcttggg gagtcgaaga ctgccttgna 300 ttgccggggagtcagcagtc gctttttccc ggncgaagcc tcgaactcac cancctgtct 360 ggattaattagacagcaaga cgcttgcggc ccctttggcg cgaacgaacn ncgaaaagga 420 cttgcgcggcccgtttcttt ggggggccaa taccggggcn cggggaaaac ccgnggggng 480 gccaaacccc cc492 27 500 DNA Human misc_feature (348)..(348) n is a, c, g, or t 27cgaagcgatg gaagcgcaag cttggtaggg gagcattccc acggcagaga aggtcgggcg 60acgagccggg ctggagcggt gggaaaagca aatgtaggca taagtaacga caatgcgggc 120gagaaccccg cacaccgaaa ggctaaggat tcctccgcta tgtcaatcaa cggagggtta 180gtcgggtact aaggcgttag cgaaggcgaa gcgccgatgt gaagggggtt aatattcctc 240cacttgccat gcgtgtgaat ccatgacgga gacgaagccg ggggtgcgtc ctgacggaag 300tgggcgccag caggggcggc cttcgggcca aaccgaacct caggtcanac ttccaagaaa 360agtgggtgaa acgccagcgc atggcaaccc gtaccgcaaa ccgacacagg tagccggggg 420anaacatcct aaggngctcg agagtacttt ctagagcggc cgcgggcccc atcgantttt 480ccacccgggn ggggtaccag 500 28 231 DNA Human misc_feature (18)..(18) n isa, c, g, or t 28 aagaaattcc gggcacgnag gcacgcccct ggtaattccc caggcgnacttctggggang 60 gctggaaggc ttgnagggca gaaaagggat ccgcctttgg gaggaacccaggtaaggttt 120 aagaaggaac ccaccctngg ggccaaacaa aaacttaaaa acccccccatttcntncccc 180 ccaaaaaaaa aatttttaaa aaaaattttt ngcccccggg ggcattgggg g231 29 109 DNA Human misc_feature (1)..(2) n is a, c, g, or t 29nncgaacaat angtctggag ctcgtgcgnc ctgnaggtgc gacactagtg gatccaaaga 60attcggcacg agggattaca gtcgtgagcc actgcacctg gctgcaatt 109 30 100 DNAHuman misc_feature (3)..(6) n is a, c, g, or t 30 tcnnnntntg gtntnggctntccgagnggc anngagtgan tgcccgttnn tattgancac 60 cantcantng ttgccntntgatacccnana caaaattgaa 100 31 100 DNA Human misc_feature (12)..(12) n isa, c, g, or t 31 tcgggcgggg anccctttac ctgtcnttac gatgcgcaag tagatnccngatttngtccn 60 ganggtcgnn aanttaggnt tccagcctgc gncacngcca 100 32 104 DNAHuman misc_feature (2)..(2) n is a, c, g, or t 32 cntgctntta cgatgcgcaaggtagtnccg tgantttagt ccgtgatgtg tcgaaanatt 60 agnnttncag ccngnnnnantgccattttn gctctnnnga gaaa 104 33 102 DNA Human misc_feature (5)..(5) nis a, c, g, or t 33 tgggntggcc cngcttaact tttgcccncg anctcggngttcgnacaggg gcgaagnaaa 60 ccgccaantt ttttcnaacc cnacttgttt tnggttttag tt102 34 100 DNA Human misc_feature (3)..(3) n is a, c, g, or t 34agnacgcctt tacagcttta ngatgcnnga gagagtancg gatttgnccn tgntggtgga 60naaattaggg ttncagcntg tgnantgcca ttttcgntaa 100 35 100 DNA Humanmisc_feature (21)..(22) n is a, c, g, or t 35 cacgatagca tcagacggcgnncttggngc cnttttgccc gctggtcaca ggacaacgca 60 tttcncnntn tggtgtncggctntcacgca tnggcgcgag 100 36 153 DNA Human misc_feature (4)..(4) n is a,c, g, or t 36 tggngccntt ttgcccgctg gtcacaggna aacgcatttc acnntntggtgttcggntnt 60 cacgcacggc agcgagtgca atgnccgatt cattcttnaa cgacgcacacacccngnngc 120 cctgtgaaac ccataaacag tgggaaatgg tgc 153 37 151 DNA Humanmisc_feature (7)..(7) n is a, c, g, or t 37 gcgcgcntgn aggccccgacactagtggat ccaaagtatt ttggcacgag ctnagttcga 60 ngatnnagac cncnnatcacctaatacanc catnactcan atgactnttt gtgcgccttt 120 tatcanatgc atagcctatcnaaaacatca c 151 38 100 DNA Human misc_feature (2)..(2) n is a, c, g, ort 38 gngcgcttgn aggccgacac taggggatcc aaagaattcg gcacgagctc gtgccgaatt60 ngncacgagt tnggctgcnt ctttatacaa cttttcttca 100 39 100 DNA Humanmisc_feature (5)..(5) n is a, c, g, or t 39 aaagngnntn ctggnnttangcanttaacc caggcactgg ggcgctgaac agctactcag 60 ctgcttaagt ngtcccactggtccagacca gcgacccagc 100 40 102 DNA Human misc_feature (80)..(80) n isa, c, g, or t 40 ttcccccagg atctttctta tatctatcag atctaggtga aaggattactgtcttgtagg 60 tgtcctgaag gacaagccgn ttcgtttgaa nctgtgaaat ac 102 41 325DNA Human 41 ttcggcacga ggagaagaga ggagccgtca gaacatatgg gggatgtgttcaagaagcag 60 atttgtggtc ggaagctttg caaagagggg acctgggtct gagtgacatgcgtggccact 120 ggtgctcctg cgtttggact gtgcaggcct ctcctatgct gatgcgtctccccactcctg 180 agctaatttc tgctctgctc cttctgtgac atgtggcagc gtgggaaatagccactgtcc 240 cctgtccctg ctgttcctgg tgtcacccag caccaggcca ctctgggagccagggcagat 300 ggtcctccct gtggtcctgg cctct 325 42 103 DNA Humanmisc_feature (14)..(14) n is a, c, g, or t 42 gtggcccaag gggnactgaaggggccctcc ntaagnggag gggttgggga gtaaggcctg 60 ggnaggaccc tgntgactcggggggcggga gcngggancc agg 103 43 221 DNA Human 43 catattttga aatacttttctcccaaactg ggtttattag cgtgtaccct gcttttccac 60 tttaaaaatt tatgccatatgtccagcttc cagtcagtgc ttctggttag catgaggata 120 actagatttt actgtagatggtagataaaa gtccagtgaa aagcaaagat gtgtaatgtt 180 ttggtagcct cagtgctcttatcccaagta aaagcaaagt t 221 44 100 DNA Human misc_feature (2)..(2) n isa, c, g, or t 44 anagagatca ntgatttatt gctgggnncc tgtntganng ntctaaggnntgaagattat 60 nncattnngc aagcgnacnn gcgcngccna gcngaccagg 100 45 106 DNAHuman misc_feature (8)..(8) n is a, c, g, or t 45 atatttcngg agcttgcagcggcnacacta ggnnactaaa agaattnnag aaagaggnct 60 atnggacnag nanacangaaacctgcanac ttggnngctt ggaagt 106 46 100 DNA Human misc_feature(74)..(74) n is a, c, g, or t 46 gatgtggaga tgcttgatag gttactgggcggcaatccag gagttgatga agcgcatatg 60 cgaacatttc acgngcatat tgcggtgcaagggcttactg 100 47 101 DNA Human misc_feature (7)..(8) n is a, c, g, or t47 ccccccnncc cttcttntcc ccnaaagaat aanataagaa tngctannga gnaancgacn 60anggtnttan nagntatatg tatntnncaa accaantann a 101 48 100 DNA Humanmisc_feature (5)..(6) n is a, c, g, or t 48 aaggnnaggc tcgttgggggaaaaaacccg ccntnncggg cncccngnaa acccncacna 60 ggggacccna aaaaccggaanaaaccnccc nagnaancca 100 49 473 DNA Human misc_feature (20)..(20) n isa, c, g, or t 49 atgagtatga aatgaaaggn tgagatgaaa tgatgatntg agatgagatgaaatgagatg 60 aaaccgagat gaaatgatga aatgatgaga tgagaccgag acgaaatgatgagatgaaat 120 gagatgagat aaaatgagat gaaatgaagt gaaatgaaat gaantcctgaaattgacntg 180 agatgaactg agataaaatg ntgagatgaa ntgatgagaa gaaatgagatgaaatgagat 240 gagatgatga gatgaaaaat gctgagatga aacntgatga gatgaaatgatgagatgaat 300 tgaantgaaa tgaaataatg aaataatgac ctgagatgan atgaantgatgaactgatga 360 actaatgaaa tgaaaatgaa atgganntga tgagatgaga agaantgctgagatgagata 420 aaatgagatg aantgatgag atgaantgaa atgctgagat gagatgagatgaa 473 50 453 DNA Human misc_feature (5)..(6) n is a, c, g, or t 50ttccnnagct gtnacganac antcttgaat tgaaattgna cacanctngt gtgnagccct 60gatanggccn gnaagcaatn tanaggatan ccgnangnta tngnaacaca ttncncnagc 120ntntncanca gctgatgcag gncncctatg atgcgattan ggactacgac tatnnctcan 180ngtctnaaca gncgcgangg ctgantacta aaagnacaca aanntgtgca ccnncatnac 240tcncgttgac tgnacantgt agacctgnaa tacctggctn aaaggggtct nactgncatn 300agagntgnag ntgcccctnc antagngnga gctnnaanng gcctgtnttt gntttacntc 360ntcgganagg cgatgccatt anagacccna gaacncattg gtgatatacn ctnnaccngg 420agggnttaca ttgggnaatg atnattatgg ggg 453 51 542 DNA Human misc_feature(19)..(19) n is a, c, g, or t 51 caactgtgag caaggaatnc cattaaatgccattgtatat tcattgatca gtgaaatcnc 60 atctgggtca cagtggcatc tatgttnacagtataaatcc ctgtggctat gaatgaaang 120 cttgtttaga cttgcatctg cacatagaagtagggatttc atgctgttat cagcctaatt 180 ttagcctata gaatttcaag ttngctagaggtttngctct ccatggtata agtttagcaa 240 gaaaagtcat ttgtctgctg ctctagcaggttanaatgtg gaagtatagt gtgcanagtt 300 ttaatccgna tatgttatta aaacatatacatcattttat atcatacatc tgnaataaat 360 attcaaaatt aaatagtgat ttgggattgattacatctta ttactagctg taataaatga 420 cctcnnngat ngtttaaaat tgttttcctcncatataata aaaatacctn angcatanat 480 cgattgtcca aaaattgaat atatatacacacctcttcca ttagaactaa atatgtggaa 540 tg 542 52 733 DNA Humanmisc_feature (13)..(14) n is a, c, g, or t 52 atatgacctg cgnncanacncnctaanang ngactngtta aanacnttcc gtggaatnna 60 ctcagactgc aaantgtnatnctgncnnan nntgnngact gtccngncng atttnnngcn 120 tgnaatacta ttgcctcttatatacacnac caannntgcg aagggcnann nnacctttnc 180 cantnnnctg gggncccacnnnngngaact gagagtggat cttgtgtacc tgacnnacca 240 gntntnnagn agggcgctcactctgattgg tgcaccatgg ttacacagtg tgtgcaaaga 300 ccngnctatc tcactgangatgattgncag ngccnntggg tggcacnang ggnactgatg 360 ancancactg accctgccgacgccagangc cgcanatccg gagantncat gngacnatat 420 aggttaccnc cttcnaccgggcancaatct gcttctatgg tgaatgcaga ccatntagaa 480 ntctntcnct ataggcatgattttnnncag tgcgtcagcc ttganaanga ancnnacttt 540 tgntagatga nnngntgctcncccttgngg ctnacaaatt ccancaccnt tggtggcngc 600 agccnttaag ancacttnttttgggttgcg ctnttggatg aattacnaat agnntgtttt 660 gttncaaggc ccttctgcnaaatatgaana aaagngcnct tagctttttg ngggaactgn 720 actggaaatt ttg 733 53100 DNA Human misc_feature (13)..(13) n is a, c, g, or t 53 gatcagacaagancntggtc cacagcggga cgagagntct cnannctgcn ggggagnnnc 60 caagtacgcnagcnctgaan ctaaagcaag caagaaaaag 100 54 515 DNA Human 54 atatggcaaggataacccct atacttctgc ataatgaatt aactaaaata acttgcaagg 60 agagccaagctaaacccccg ataccgacga gtaccagaac aggtaagcac cccgtctatg 120 tagatatgggaagattatag gaggcgacaa ctaccgagcc tggtgatagc tggtgtccaa 180 gaagagtcttagttcattta tttggcccag aaccctctaa tccccttgta atttatgtca 240 agaggaacagctctttggac actggaaaac cgtgagagag taagatttac acccttaggg 300 gcctaatagcagccaccatt aagaaagcgt tcgctccaca cccactacct aaaaatcgaa 360 tataactgactcctcacacc caattggcca atcattcccc tataaaagaa ctatgttagt 420 ataagtaacctgaaaacatt ctcctctgca taagccctgc gttggattat atcctgcact 480 gacaattaactgccccaata tctacaatcc aaccc 515 55 176 DNA Human misc_feature (5)..(5) nis a, c, g, or t 55 tgttnaggat caaattataa tattgaaata anaacagctnacatttatat agcatgtttn 60 cntatctcaa ctaatnataa atgggaaaat gggcaactgggcaggcngaa cccagaggga 120 agcctgccct cattagacca agacagcaag gtttnccctggtcactagat gaaatt 176 56 317 DNA Human misc_feature (4)..(4) n is a, c,g, or t 56 cagnagtgat gttgcaatat ctggaactag caaaggatac tgatgagaaaacgtggaatc 60 atgtgggatg tgacctccta ggactcacct tgcacagctg ggtgcagcagggataggtaa 120 ggatttgggg tttagaggta caattgcctt tttatggtta gagaaaggtcctggggctgg 180 agggagcctg acgatctgct ctgtgtgcaa ggggagagtt aactctgcacgcaagagcct 240 gcttaaaggg ctgtgtcagt tctattgtaa acaccaactt aaagtggtggatgctggcag 300 acattgttat tgccatt 317 57 209 DNA Human 57 ctcatacacctgtggctact gttttctaca gagtgccaaa actattcgag agaataggct 60 ctggactggacactgtatac ccacatgcaa gatgaagttg gccccttaca tcctatacgc 120 aggagaattgcgtcatttaa agcctgttga cgcttttctc ccgcagacga atggaaagat 180 taattgggagtgggggctga aacaattcg 209 58 262 DNA Human 58 aattttgctg ttacatggtggctcaactga gtcccatact ttgaaggccg ggagttaatc 60 acctggtcac cgagttgcgaaccagcctcc aatatgtgga accctgtact ctctaaaaat 120 caaatcaccg gcatggagattgcgcctgtg gtcccaaaat actcgggctg ggacacgatg 180 agttgcttgg cccaaggaaggagggttgta tggctgatca cactggtccg cctgggtgac 240 agagcgagac tccatctcta at262 59 430 DNA Human 59 gtcagtttat ttctgactag ggatattttc tttccatttagaaaagaaga aaaaaaaaaa 60 aaacctttat tgtcttacag gggggaacta gcgcggggctgaataaaacc tttggccctt 120 cccgggggag gggtatccgg tttataaacc ccaagggtattttcttagca aaatacttaa 180 aaccggccgg ggtttttata caaactggga acccacttttgaaaaatttt ggccttttga 240 tctgggatgg gaatatgagt ttttatacat ttcattttctttttgggcaa aggcccggtt 300 aagtattccc ccccgggggg cctttacaaa aagggcggttttaaaagctt ttgggccccc 360 ctagggaatt gttttaacac ctaaaaaccc ctgcttcccttaaaggggcg ttctttaatt 420 tgggggcggc 430 60 350 DNA Human 60 aaacctctctaactatatat cacaataacc tgcgcataag atttacgctc cgatcttttc 60 atcctactagcttggaggat ttgaaccgat tatgaatacg caatactccc ggtcctcatg 120 tatcatgtgtaagcccatct cctgggaggg ctaacatact accatctcca aggagaggca 180 tgattccgaatcacccacag acagctcgat caccatacgt atcacccaac atatatacct 240 tctaagacttgctagaaaca accaccacat ttgatgctta atcaccactc tgacgcgcat 300 taaagtgaggggactctcct aatttctgta agttgatttt tgcattctga 350 61 515 DNA Human 61cacataaatt ctccataagt taattagtga ttttaacatg atctcaatat aaacatagca 60cactttcttt gagaattcaa catattgcaa gttaaaattt tcatagacta cacaagaaag 120aataatcagg caaatcctta agaataaggg caattaagga tgactagccc tacaagattt 180taaaaaggat tcattagttt aaaaaatgtg atgtagatac atgaataaaa taaaatcttg 240aagtagatcc aaatatacat ggtcagattg aatacaataa agatggcatc gtagcagtgg 300agaaaagaag aattatttca taaaccttgt tggaatggct aggcaatcat ctggaaaaaa 360atgaagttga ataataaaaa tatattctac actagcacaa attataaata aagcagtgat 420ttaaatgaga aaaattaaat cataatgatt tcaaagataa cataggataa tttctttata 480gtcttctaaa atatatgact ttatgaattc tgact 515 62 611 DNA Human 62caagtacttt accaactaag ccaatcttgt ccccagccag gcatttctat acaaagggcc 60aagactttgg ttttataaat aaggaggtat atataaatta tatatatttc tgagctgagt 120aataatccac cagatacaag tttgcatcaa cttctgtgaa atattttttt tcctttttgt 180tgggcatttt tatggtctaa atatagaatg accaatgcct ctagaacaaa cttgacctgg 240tcagtgttat caagaagcag actgtttctt actttctttg tatttcctta cttatttaaa 300tttgttaaaa ttgatatatt gatatataaa acttcttttg ccagtgttgg tggcacacgc 360ctttaatccc agcacttagg aggcagaggc agggtggatt tctgaatttg agggcaggct 420agtctacaga gcaagttcca ggtcagccaa ggctatatat agaaactctg gcatgaaaaa 480ccaaccaaac caaaccaaac caaaccagac cagaccagac cagaccagac caaaccaaac 540caaaccagac taaaccaaac caaaccagac cagaccagac cagaccagac cagaccagac 600cagaccaaac t 611 63 291 DNA Human 63 ccgagagatt ggccactgct taaactcatgcagctcctac tgttcttcaa ttaatgcctt 60 taatgcgaat atacttcctc ttctttttgcatggtcttgc ccagcctctg caatactgat 120 gaacacatgc tgaagatcat ctaactcaatatggcgcata tttctatgtc ttgctgccca 180 ggacatagga caacttcgtc gctcactagttctaacatat taatgctggc gtaggtggag 240 aactactgca catatactct tactcggaggctgaggcacg aggatcactt g 291 64 309 DNA Human 64 gccagatgcc gtgtttcctcgatgaactct ttacatcatt ggctattcag tggagtgttt 60 cattatcacc tctcactctcgcgtgttacc taactctccc tcgcagggga aatcactcca 120 tatatttcaa atgtcttgctaacagtggtt actttgctct atccttagct atacgtctcg 180 aggcacattg ttcctctatgccccgctacg ctttgcccta gagctcggcg gtatctatat 240 cttaactgcc ctcttgatccttacgtgccg gagaaggtgg aggcagaaat tttgtcaaat 300 ctgattaga 309 65 278 DNAHuman 65 tagaatggaa tggagtcgaa tgtgatggaa tggacgcgaa tggaatggaatggactcgaa 60 tggaataaag tggaatagac tcgaatggaa tggaatgcaa tggaatggactcgaatggaa 120 agggatggaa tggactcgaa gggaatggaa tggaatggat tcgaatggaaaggaatggaa 180 tggactcaaa aggaatggaa tggaatggac tcaaatggaa tggactcgaattgaatgaaa 240 tgtaatggaa tagactcgaa tggaatggaa cgaaattt 278 66 142 DNAHuman 66 agttctcctt aggttaatta atggaatgca atcccaatga aaatgtcaccaaagttgttt 60 tttttttaac tgtaggaggt ttataataat gctcatatgg aaaaataaaacatgtaaaaa 120 atagctagta aactccccct gt 142 67 286 DNA Human 67atatctgcca tcctcatcgg ccaatcgtgt tattttgatg acgaatgctt cggagattgg 60aaagatgatc tcctcatgct tccatgcact gcgagtagaa gacatactga gcatagtgtg 120attattttcc caacaaattg gcattcatag atagaataag ctgactaaga ctacttagcc 180ccacattttt ttctacttgc tccaatagca ctaacaaata ggaagctctt gcttgctccc 240caaagctcca tttccttgaa agcagaagtg taatattact tcttag 286 68 179 DNA Human68 atctactttt tattcttttg ataaatgttt atgaaatata aaatactgaa aattagaaag 60tagaagtcat tattttatta taaaacatgt ggattagata ttttcattta tgtgattaaa 120ctttctaaac aaagattata tgaattatct taaagattta aaaagtaatt aagttaaat 179 69390 DNA Human misc_feature (356)..(356) n is a, c, g, or t 69 cagataagactattaagaca gataagagcc aaatcatgta gagcctcaga ggtttttgat 60 cttcagtctaagaacgtaaa tccatggaag aattttaagc aggggtgtgc cttgaccaca 120 ttttgaattctaaactgtct ctgggtgggt gtgggtgcca ccaagagcat gtgttcatgt 180 agggagactggttttttaca gttgtctatg agagagatga cagttgcctg gattatggtg 240 gtgacattggagataagcag gtagacagat tctcagtgta ttaggagaga aaaatcaata 300 ggaaatttaaaataaataat taactgtggc cataggagga aggagtcttt gggttnggtt 360 ctcaatttctgcatgagaaa aaaggtggac 390 70 481 DNA Human misc_feature (26)..(26) n isa, c, g, or t 70 atgatgaaat gatgagatga aatgcntgag atgagatgtg atgaaatgatgatatgaaat 60 gatgacataa aatgagatga aatgagatgt aatgatggaa tgagatgagatgaaatgaga 120 tgaaatgata gatgagataa aatgatgata tgaaatgatg agatgaatgatgagatgatg 180 agatgaatga tgaaatgaaa tgatgagatg agatgatgaa atgaaatggtgagatgaaat 240 gatgagatga aatgaaatag tgaaatgaaa ttgaaataaa atcgaaatgagagatgaaat 300 gatgagatga tgaaattgat gaaatgatga gatgtgatga gatgaaatgatgagatgaga 360 tgagatgaca tgaaataatg aaatgaaatt gaaatgagat aagatacgagctgagatgca 420 atgagatgaa atgatgagat gaaatgaaat agtgaaatga aattgaaataaaatcgaaat 480 g 481 71 125 DNA Human misc_feature (5)..(5) n is a, c,g, or t 71 cggtngcaat tgggggccnc atacgcgcng acgagtantg gncangctncttgactacac 60 ngacgcgccg tacaggntna attatggnan cttacatggn aaaggggcanctcaatgtcc 120 cacag 125 72 473 DNA Human misc_feature (151)..(151) n isa, c, g, or t 72 gaaatgaaat aatgaaatga gatgaaataa cgaaataaaa ttgaaatgagatgagaggaa 60 atgagatgaa atgttgaaaa gaaaggagga aatgatgagg tgagatgaaatgatgagatg 120 aaatgaatct gagatgaaat gagatgaaaa ntgatacgaa aaatgatataaaaaatatga 180 cctgagatga aatgagatga aaaatgatac gaaaaatgat ataaaaaatatgacatgaaa 240 tgaaatgaga tgatatgaaa tgacataatg aaatgatgaa ttgatgatattgaaatgaaa 300 ttgaaagatg agatgaaatg atgagatgaa atgaaatgtt gaaatgatgaagagatgtga 360 catgaaatga gctgaaatga gatgaaatga aatgagatta aatgatgagatgaaaaatga 420 tgagatgaaa aatgagatga gatgatgaga tgagatgaga tgaattgagatga 473 73 500 DNA Human misc_feature (7)..(7) n is a, c, g, or t 73aatgagnatg aaaagnatga aatgatgaga tgaaatgaaa tgatgagatg aaatgaggtg 60aaatgaaatt agatgaaatg taatgagatg aaatgaaatg acctaatgaa atgaaataat 120gaaatgagat gaaataaaat aatgaaatga tgaaataatg aaatgaaaat gagatggaaa 180tgatgagatg agaagaaatg atgagatgaa atgatgaaat gatgagatga ganaaaatga 240gatgaaatga tgagatgaga tgaaatatga tgagttgaaa tgacataatg aatgaaatga 300tgaaatggaa taatgaaatg gaaatgatga gctgagatgc aatgagttga aatgagatga 360aatgatgaaa tgatgagatg aaatgatgaa atgaaataat gaaatgagat gaaataaaat 420aatgaaatga tgaaataatg aaatgaaaat gaaatggaaa tgatgagatg agaagaaatg 480atgagatgaa atgatgaaat 500 74 299 DNA Human misc_feature (31)..(32) n isa, c, g, or t 74 ggaaatcctg aagtggaaat gatgagctga nntgcaatga gttgaaatgagatgaancga 60 tgaaatgatg agatgaaatg atgagatgag atgtgatgaa atgatgatatgaaatgatga 120 cataaaatga gatgaaatga gatgtaatga tggaatgaga tgagatgaaatgagatgaaa 180 tgatagatga gataaaatga tgatatgaaa tgatgagatg aatgatgagatgatgagatg 240 aatgatgaaa tgaaatgatg agatgagatg atgaaatgaa atggtgagatgaaatgatg 299 75 155 DNA Human 75 agtgaaatga aattgaaata aaatcgaaatgagatgagat gaaatgatga gatgatgaaa 60 taaaatgatg aaatgatgag gtgatgagatgaaatgatga gatgaaatga tgagatgaga 120 tgagatgaca tgaaataatg aaacgaaattgaaat 155 76 367 DNA Human misc_feature (11)..(11) n is a, c, g, or t 76atagcaaaag ngggtaaaac ccctgagttt gcganannag tantcttgta ggggcnaact 60ctacttnaga ngaantcctc gcaaaatcct tgaatcaccg cttcagtgca gtgatatcac 120cgccatgaaa tttctgctcg attagcttac gttgtttgga tagaggccaa acaaggctgt 180tatcggtacg aggaatggat gttcgatttc gtagaatacg cctgagagac ggcgaatact 240ctcacgagag gcagcaggcg cgtaaattac ccaattacaa caagtagagg tagcgaagga 300aaatatgagg ggtggcaagg ttttgcctgt tacattctca aatggaagca aattagatat 360gtcattg 367 77 257 DNA Human misc_feature (6)..(6) n is a, c, g, or t 77actagnacag naattttagc taagtggagt ttgagttaag tggagatgtg agaccatctc 60atagaaatca ttatttctgt gggatggata attgggccaa attgtaaaat attttaacta 120tcagtgtttg gggtttattt ttaaaagaat agggtgccac cagatgttct ttagtggagg 180agaaatgagg ccagagtgac tgcctagaaa attaagttgg taaattaatc acttttttct 240aggtcctttc ttagtct 257 78 373 DNA Human misc_feature (11)..(11) n is a,c, g, or t 78 ctttaaaaac ntgttagacn aacnttaaaa nttacccntt ttcctgaactgantcctggg 60 nntaantaaa aagggtgaag aannttactt cncttggtcc taaaaaacnttttcntcagt 120 tattaccaaa atatttggac cattantaaa gantagggcc aacccnaatttttcttgaaa 180 tttccgttaa atagccgtta aatgttttta cccatttcat attggataccttaaattata 240 ataatggatt ttattgttaa attgtgtgtg tgtggtgtgt atgccctgtcttttctcctc 300 taccattatt gtcactttat gtttggaacc ccctttaccc ttccttaaaggaaaaaaagg 360 gcccggggtt ttt 373 79 128 DNA Human misc_feature(10)..(10) n is a, c, g, or t 79 tcctagtaan ctggtttacn ctgaaagannaagangcctc ccctgttcnc tgaaatacca 60 ccttgatgtt caagtattta agaccctatgcnaatatttt ttaccttttc taataaacca 120 tgtttgtt 128 80 213 DNA Humanmisc_feature (9)..(9) n is a, c, g, or t 80 cccattggna cagacccccaaaatgggtac attttttagg aaaccaggac ctttccaagg 60 ggccaggcct tccctttaaaaaaaaatnac cgtttttngg gggangnaac ctttaaaagg 120 ggaaaanaaa tcctttttaaanggaantcc aagggaagga ncctgnncaa nacttccccn 180 ccaataaaaa aaaccnttttggaaangggg aaa 213 81 443 DNA Human misc_feature (22)..(22) n is a, c,g, or t 81 gaaatgagat gaaaccatga gnatgaaatg aannaatgnc atgcaaatgatgagatgaaa 60 tgatgaaatg agatgagatg agaagaaatg acttgatgag atgagataaaatgatgaaat 120 gaaatgaagt gaaatgaaat tgaaatgaga tgagatgaaa tgagataaaatgatgagatg 180 aaatgagaag aaatgagatg aaatgatgaa atgatgagat gagatgaaaaatgatgggat 240 gagaaatgag atgaaatgat gggatgaaat gaaatgaaat aatgaaataatgaaatgaaa 300 tgaattgata atattgaagt gaaattgaaa gatgagattg gatgaaatgatgagatgaaa 360 tgaaatgttg aaatgaaatg aagagatgta acatgaaatg agctgaaatgatgagatgaa 420 atgaaatgaa atgagattaa atg 443 82 442 DNA Humanmisc_feature (13)..(13) n is a, c, g, or t 82 tggcccggga acntcnaactgcccatcctg ganttttggg ggggannctt taaaaaacct 60 gacctctgaa tgtattantganncaagtga tagccaagat attttgaaga aaaatagata 120 ntagggacct gctctataagcccatcataa tttattatga agttataaca agtaaaacag 180 taaggtattt ggcatggaatagagaaccca gaaacagacc caatgcatgg gtacaggata 240 taacacaggg aaatgagggacaatatatgg ttctgggata attatttata tggggaaaat 300 aaagaaattg gatccctacctcacacatac aaaaaaaatc ataattgaat taaaaacttg 360 catgtgaaag gaaagactttaaaacattta gaaaaagtat tggaggctat gatcttgggg 420 taggaaagca tttctttttt tt442 83 135 DNA Human misc_feature (8)..(8) n is a, c, g, or t 83gtctaacnta aaaagtaaag aaagtaaagt aaaggnttga aggaaggaag gaaggaagga 60aggagggaaa agaaagaaag gaaggaagga aggaaaagaa agaaagaaag gaaggaagga 120aggaaggaag gaagg 135 84 346 DNA Human misc_feature (30)..(30) n is a, c,g, or t 84 ggaggaggaa gagtgatgag ttctctaatn acttggttgg attagccttagagttatcgg 60 gagttgcctt ctgtaagtgc ccctactatc aaggtttcat ggaaaatctaggcaaggcag 120 aacttcctca gaaggacaag agacaaagaa gtgggggagg ccctcctatccatagctgag 180 agggtttatt ctttgtggtt ctgctgtcag agcctttgga tgtctgatctgagatggagc 240 aaccccagct agacagaact ttgtagattt tggggggttt aaaaggcctcaagcaaattc 300 taaaactttc tttgaacccc ctggcatagg ctcagtttcc ctgact 346 85100 DNA Human 85 acaaaaagcc cctttaaact tgggcccgct cgaggtcgtt tcgactgggccgagacttcc 60 gaaaagaaaa tggttttttt tgccgaaatc aaccgggtaa 100 86 201 DNAHuman 86 ttcataacat cgtcattttg ggttatgcga aatacaaatt taaatctttgtgaaatgaaa 60 gaaaagagga agaaacgctt tttaggagtt aaggattaaa gtaaaaattattttgacata 120 attacctctt tttgtgacca ctcttaaagg ccaggaacat atttggagaagcctagttgt 180 atgtaacagt gtggggtttc a 201 87 531 DNA Human 87tatagcgggc gttataaaca taccacttcc cggtacaacg gatttcaagg ttaggggtgc 60aacccagaac gaacgcgtta agtgcgcgtt atcttcctag gatagagtcg gtgacgggaa 120tcttttaccc cggcactcgg gtccaccctc gcggcaccag aggtattctc cggcgagtcg 180ttaaccatcg caatcgccga ccgagtttaa ggaccactcc ccacctttct cattagttaa 240ggagaacgct actttacccc atagacggag aaatcgctac tcaactacca ggcgcgcgcc 300gtcgagtccc tcttcctctc tttatgcatt tagagcgctt tcgtaagagt tttccctaga 360ttcttctaag cgtagcgcgt ctactccaat gttttcgtta atccagcccg aactaacgcc 420gcggaggagt cgatccgtct actcctatcc cgtcggctcg gatttactac aggagctaag 480aaaacaaaaa gtaccagccc taaaggaaag tcaaaggacg cccgtaaaaa a 531 88 530 DNAHuman 88 aatctcgatc gcaaacatac ggcactctcc ctcttgccgc ggttttcgtccagcgctttc 60 cattcggtcc agtgcctcgc cctattagcc cttaagccca ccgtttctaaaactcccaga 120 acagccaaac cggtccgccc aaggcctccg tcgttttata atatattccgtttacgtata 180 aggaacgaac cccccttcat taccacggtc ccgcgtccgc ctccttctccattcgcaaca 240 gttctattcc tttcagcctc ccgtacctgc ttccagaaca tcgcaccgccatagtcgaaa 300 gatagcaaag attacccagc ttctattcct cgccccagag ccgagtaaatcgaagtttat 360 agaggcggaa tccaaccatt caagagttat aacaagttat cggcactcgggggatcagaa 420 tataaactta atgtcccctt tattctcccg gacgcccctt ttaaccacttcttcctatct 480 ttcgctaaca agccattgac ggcgctttgc cgcgcgggcc catctcgcgt530 89 332 DNA Human misc_feature (37)..(37) n is a, c, g, or t 89ccatttatgg gccggggata tacccacatg gtacagnaca ttacatnttt atggcaccat 60ttccaccggc ctggttttgg tttttccata attaattaac cagggggncc anttaaaaaa 120aattaaggna aggnttaaaa aatttaacca anggggggtt taaagggntt ttttttttta 180aaaaaaaagg ttaaancccc cccttttttt ttgggttggg gtgggaaaat tttgggaanc 240cttaaccccc gggtttttgg gtttttttgg ccaaaacccc ccggaaaaaa attaaaaaaa 300ggaccggttt ccattttaat gggtattggg aa 332 90 185 DNA Human 90 actgctataatgcaggggaa catgttctca gggtcatcct gaggggttgt gtcatggggc 60 cggtggtaactattaaaaca taagtttaat cggtatttaa aattttaaaa tcaaaaaaaa 120 taaaatatatgcaaccctcc attccaagga agtatgatgt tactagatta tctgaaaatt 180 ctcct 185 91365 DNA Human misc_feature (326)..(326) n is a, c, g, or t 91 ccagagagccacaaatgacc aaaatatttt gagatgaaca tgctcgtaga aggtagctga 60 ctagggggtacttgaaaatg ctagaccagg ataactccta agtgtatatc cttggcagac 120 tcgttatgctttccaatcct gcttgcaata taagacacaa agtcagaata aagctcaaga 180 aaacagaacgtgcaggccat caagcgcaga gcctgctcat tggacaaccg caaagagtag 240 taagtgctgccgctattcac acttagaaaa ggagaaccac ggggaaaaac caaattaatg 300 gggctgctttttgtcactct ggcatnagag aattgtgnng aaantttaac ttttgtaagc 360 ttgta 365 92113 DNA Human misc_feature (32)..(32) n is a, c, g, or t 92 acttgaccttatggatgatg ctgcggagtg cntngtaagt gtttcatgat attccttaag 60 aagtcaggatagtagttttc attccttaga tggtacaagt gttgagacaa atg 113 93 210 DNA Human 93gttttaggga aatttgccag ttttatgttt taatattttt ggaaggaaaa ctgaaaggta 60atgaaaatgt tactgttgga ttaaaaaaca aattaagtcc aaatagtgat taggcaagtt 120ggtgaggtag ggggttgctg caagagcgga agttgaaaga tcttggaaaa attaaagaaa 180cttcatagaa ccccatctct acaccaaaaa 210 94 506 DNA Human misc_feature(5)..(5) n is a, c, g, or t 94 ttggnggggg ggcgagatcc tactngagacccttgatnnt gggnanggac cgaagatcna 60 ttaganaccn atgngatggn cnnncnaaannnttaaagtg agagtccatc tnngaanaaa 120 atgggnaant ttnnnngggg ggggggaaaaancccnnggg tnannggggg cccngggntt 180 naaannnggn nctngggggg ggaaanttttggcccccccc cgggggnttt ncctnaaaaa 240 aaanccnttt naaanacngn nanaattttnccnnnncggg gaggngngga nntttttttt 300 tnaannagcc ntttttgnna naaaaannntggnccccccc ctattccnng gnttttngga 360 ccnttnnanc ntgggnnttt ttagnccttnaaaaaaangc naatnttaag gtaaaaattn 420 ggggggggng ggggggnggn gnntttttttttntnnggag gggttttttt ccnncgnggg 480 ngaaagnntg gggcnnnctn cngccn 506 95400 DNA Human misc_feature (11)..(11) n is a, c, g, or t 95 catgaaggaanaagcctgta ctanctgccg gtatccatgn taatctgngg ngatgtcagc 60 agacccagctnagcagatan ctncatttct ntctnaagnc ctttggtctg naggnngnca 120 ntnnanctncngntnaacat cacagctnct ccnagcatca ccctgctagn tancngnggg 180 ttttctcttatntgnngncn naacatctgc nngctctgnt annaanaatt ncataccgcn 240 canngtctntgacgntgtga tgcatacgnt tgggcagagn gancaatang tgngcatatg 300 cgtgccttacncaaggatac ggangngctt gaaattgatg ngaccaanan tttnngtacg 360 gtaagtnacccaaccacttc tgnnttcact ntaagagncn 400 96 800 DNA Human misc_feature(171)..(171) n is a, c, g, or t 96 gagatgaatg atgaaatgat gagatgagatgatgaaatga aatggtgaga tgaactgatg 60 aaatgaaatg aaataatgaa atgaaattgaaataaaattg aaatgagatg agatgaaatg 120 atgagatgat gaaataaaat gatgaaatgagatgtgatga gatgaaatga ngagatgaaa 180 tgatgagatg agatgacatg aaataaatgaaataatgaaa tcgaaatgag atgagaagat 240 acgagatgag atgaaatgat gagatgaaatgatgaaatga gataagatga aaagagttga 300 tgagatgatg agatgaaatg agatgaaaagagatgaaatg agatgaaatg aaatgatgag 360 atgaaatgag gtgaaatgaa attagatgaaacgtaatgag atgaaatgac ataatgaaat 420 gaaaaaatga aatgaaataa tgaaatgaggtgaaattaaa tgagatgatg aaattaaatg 480 atgaaatgaa ataatgaaat ggaaatgaaatggaaatgat gagatgaatg atgagatgaa 540 atgatgagat gagatgtatt gatgagaggaaatgatgaga tgtaatgaaa tgagatgaaa 600 tgaatgagat gaaatggaat antggaanggaaattgattg gngatttgag atgaaatgag 660 ntaaatgnga tgaattaatg atgagatgaaatgntgaatg ccggggtgnn tgagatgaat 720 tgagttgaac cctgngatga atgaagattgnntgaatggt ggntgaatgt tgaatggntg 780 gntggnanaa tgcctgtngg 800 97 334DNA Human 97 gatgaattga aatgaaatga aataatgaaa taatgaaatg agatgaaatgaaaagaaatg 60 atgaaatgat attgaaatga aattgaaaga tgagatgatg agatgaaatggtgaaatgtt 120 gaaatgaaat gatgaaatga atagatgtga catgaaatga gctgaaatgatgagatcaaa 180 tgaaatgaaa tgagattaaa tgatgagatg aaaactgatg aaaacttaaatgatgaaata 240 atgaaatgaa aatgaaatgg aaatgatgag atgagaagaa atgatgagatgagatgagat 300 aaaatgagat gaaatgatga gatgaaatga tgag 334 98 100 DNAHuman misc_feature (17)..(17) n is a, c, g, or t 98 ttcaggccgtctgcttntac atatactatc gagaatggtg ctgtgcactc ataacaccgt 60 tgcttggtagacgcttttga acccttcagc gctgaaagta 100 99 500 DNA Human misc_feature(8)..(8) n is a, c, g, or t 99 cccgggantt cggcccttat ggcccggggaaatgatgaga tgaaatgatg aaatgagata 60 agatgaaaag agttgatgag atgatgagatgaaatgagat gaaaagagat gaaatgagat 120 gaaatgaaat gatgagatga aatgaggtgaaatgaaatta gatgaaacgt aatgagatga 180 aatgacctaa tgaaatgaaa aaatgaaatgaaataatgaa atgaggtgaa attaaatgag 240 atgatgaaat taaatgatga aatgaaataatgaaatggaa atgaaatgga aatgatgaga 300 tgaatgatga gatgaaatga tgagatgagatctaatgatg agaggagatg atgagatgaa 360 ntgagatgaa aagagatgaa atgagatgaaaccgaaatga tgagatgaaa tgaggtgaaa 420 tgaaattaga tgaaacgtaa tgagatgaaatgacataatg aaatgaaaaa atgaaatgaa 480 ataatgaaat gaggtgaaat 500 100 397DNA Human misc_feature (8)..(8) n is a, c, g, or t 100 cccgggangtttaagttagg gggcctgccc ctttaagcnt agtcccaccn tgaaanacac 60 tccccttgaanntctctaaa ccttaacttt ctggccnttt tgtttcagan atgcctaacc 120 ctcagggggtcttttgttct ctacgcctaa aaacttaatc tgtttggaac aattccnttt 180 cctctctgtagaaattgacc tggccatggc tcctgtgaat gatacggttg ctattatccc 240 tgaacactgtaaaaatgaac tttgaaacag ttgggtagga cccaaacaga aaatgatgta 300 tggcttggaaatagtttagc tgaacattat gctttaatat tttactggcc attgcagcac 360 aggtttagaaatttatgttc ggctttttaa agtttta 397 101 132 DNA Human misc_feature(121)..(121) n is a, c, g, or t 101 gttacctaat gttttactct cattttctttttctttattt ttcatttgta aaataggaac 60 attaattgta ctactttcaa aagaattaattgaagaaaga gagatacagg gtatctaggc 120 ngaggaagac cc 132 102 246 DNA Human102 gggggcttta gttataactg ggctaagcat aattgcgcta ccaattccat attatctcat 60ggcacttaat tttataattg atatatataa taaaaaattc aatgcagata ttgatataat 120aaaaatagat aatggtaatc caagcacgat ggtagccatc actctaattg ctttggggtt 180aacctataac ttattaagta aagtgccaga atggttcttt gacagtatta aaattaaaga 240aaacag 246 103 18 DNA Artificial Sequence forward primer of exon 1 ofinsulin gene used for quantitative RT-PCR analysis 103 gccctctggggacctgac 18 104 18 DNA Artificial Sequence reverse primer of exons 1 and2 of insulin gene used for quantitative RT-PCR analysis 104 cccacctgcaggtcctct 18 105 24 DNA Artificial Sequence forward primer of aMyHC geneused for quantitative RT-PCR analysis 105 gctggaacgt agagactccc tgct 24106 24 DNA Artificial Sequence reverse primer of aMyHC gene used forquantitative RT-PCR analysis 106 ggatccttcc agatcatcca cttg 24 107 20DNA Artificial Sequence forward primer of ANF used for quantitativeRT-PCR analysis 107 ggatttcaag aatttgctgg 20 108 20 DNA ArtificialSequence reverse primer of ANF used for quantitative RT-PCR analysis 108gcagatcgat cagaggagtc 20 109 20 DNA Artificial Sequence forward primerof APP used for quantitative RT-PCR analysis 109 ggatgcttca tgtgaacgtg20 110 19 DNA Artificial Sequence reverse primer of APP used forquantitative RT-PCR analysis 110 tcattcacac cagcacatg 19 111 21 DNAArtificial Sequence forward primer of ZFP used for quantitative RT-PCRanalysis 111 cacargagrc arggtcaacg a 21 112 22 DNA Artificial Sequencereverse primer of ZFP used for quantitative RT-PCR analysis 112ggattaaaat gaagcaccca ga 22

What is claimed is:
 1. A method of identifying one or more markers fordiabetes, wherein each of said one or more markers corresponds to a genetranscript, comprising the steps of: a) determining the level of one ormore gene transcripts expressed in blood obtained from one or moreindividuals having diabetes, wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordiabetes; and b) comparing the level of each of said one or more genetranscripts from said step a) with the level of each of said one or moregenes transcripts in blood obtained from one or more individuals nothaving diabetes, wherein those compared transcripts which displaydiffering levels in the comparison of step b) are identified as beingmarkers for diabetes.
 2. A method of identifying one or more markers fordiabetes, wherein each of said one or more markers corresponds to a genetranscript, comprising the steps of: a) determining the level of one ormore gene transcripts expressed in blood obtained from one or moreindividuals having diabetes, wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordiabetes; and b) comparing the level of each of said one or more genetranscripts from said step a) with the level of each of said one or moregenes transcripts in blood obtained from one or more individuals havingdiabetes, wherein those compared transcripts which display the samelevels in the comparison of step b) are identified as being markers fordiabetes.
 3. A method of identifying one or more markers of a stage ofdiabetes progression or regression, wherein each of said one or moremarkers corresponds to a gene transcript, comprising the steps of: a)determining the level of one or more gene transcripts expressed in bloodobtained from one or more individuals having a stage of diabetes,wherein said one or more individuals are at the same progressive orregressive stage of diabetes, and wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordetermining the stage of progression or regression of diabetes, and; b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals who are at aprogressive or regressive stage of diabetes distinct from that of saidone or more individuals of step a), wherein those compared transcriptswhich display differing levels in the comparison of step b) areidentified as being markers for the stage of progression or regressionof diabetes.
 4. A method of identifying one or more markers of a stageof diabetes progression or regression, wherein each of said one or moremarkers corresponds to a gene transcript, comprising the steps of: a)determining the level of one or more gene transcripts expressed in bloodobtained from one or more individuals having a stage of diabetes,wherein said one or more individuals are at the same progressive orregressive stage of diabetes, and wherein each of said one or moretranscripts is expressed by a gene that is a candidate marker fordetermining the stage of progression or regression of diabetes, and; b)comparing the level of each of said one or more gene transcripts fromsaid step a) with the level of each of said one or more genestranscripts in blood obtained from one or more individuals who are at aprogressive or regressive stage of diabetes identical to that of saidone or more individuals of step a), wherein those compared transcriptswhich display the same levels in the comparison of step b) areidentified as being markers for the stage of progression or regressionof diabetes.
 5. The method of any one of claims 1-4, wherein each ofsaid one or more markers identifies one or more transcripts of one ormore non immune response genes.
 6. The method of any one of claims 1-4,wherein each of said one or more markers identifies a transcript of agene expressed by non-blood tissue.
 7. The method of any one of claims1-4, wherein each of said one or more markers identifies a transcript ofa gene expressed by non-lymphoid tissue.
 8. The method of any one ofclaims 1-4, wherein said diabetes is either symptomatic or asymptomatic.9. The method of claim 8 wherein said diabetes is type II diabetes. 10.The method of any one of claims 1-4, wherein said one or more markersidentifies one or more genes selected from the group of genes listed inTable 3G.
 11. The method of any one of claims 1-4, wherein one of saidone or more markers identifies the insulin gene.
 12. A method ofdiagnosing or prognosing diabetes in an individual, comprising the stepsof: a) determining the level of one or more gene transcripts in bloodobtained from said individual, wherein said one or more gene transcriptscorresponds to said one or more markers of claim 1 and claim 2, and b)comparing the level of each of said one or more gene transcripts in saidblood according to step a) with the level of each of said one or moregene transcripts in blood from one or more individuals not havingdiabetes, wherein detecting a difference in the levels of each of saidone or more gene transcripts in the comparison of step b) is indicativeof diabetes in the individual of step a).
 13. A method of diagnosing orprognosing diabetes in an individual, comprising the steps of: a)determining the level of one or more gene transcripts in blood obtainedfrom said individual, wherein said one or more gene transcriptscorresponds to said one or more markers of claim 1 and claim 2, and b)comparing the level of each of said one or more gene transcripts in saidblood according to step a) with the level of each of said one or moregene transcripts in blood from one or more individuals having diabetes,wherein detecting the same levels of each of said one or more genetranscripts in the comparison of step b) is indicative of diabetes inthe individual of step a).
 14. A method of determining a stage ofdisease progression or regression in an individual having diabetes,comprising the steps of: a) determining the level of one or more genetranscripts in blood obtained from said individual having diabetes,wherein said one or more gene transcripts corresponds to said one ormore markers of claim 3 and claim 4, and b) comparing the level of eachif said one or more gene transcripts in said blood according to step a)with the level of each of said one or more gene transcripts in bloodobtained from one or more individuals who each have been diagnosed asbeing at the same progressive or regressive stage of diabetes, whereinthe comparison from step b) allows the determination of the stage ofdiabetes progression or regression in an individual.
 15. A method ofdiagnosing or prognosing diabetes in an individual, comprising the stepsof: a) determining the level of one or more gene transcripts expressedin blood obtained from said individual, wherein said one or more genetranscripts corresponds to said one or more markers of claim 3 and claim4, and b) comparing the level of each of said one or more genetranscripts in said blood according to step a) with the level of each ofsaid one or more gene transcripts in blood from one or more individualshaving diabetes, c) comparing the level of each of said one or more genetranscripts in said blood according to step a) with the level of each ofsaid one or more gene transcripts in blood from one or more individualsnot having diabetes d) determining whether the level of said one or moregene transcripts of step a) are characterized as classifying with thelevels of said transcripts in step b) as compared with levels of saidtranscripts in step c), wherein said determination is indicative of saidindividual of step a) having diabetes.
 16. A method of determining astage of disease progression or regression of diabetes in an individualhaving diabetes, comprising the steps of: a) determining the level ofone or more gene transcripts expressed in blood obtained from saidindividual having diabetes, wherein said one or more gene transcriptscorresponds to the markers of claim 3 and claim 4, and b) comparing thelevel of each of said one or more gene transcripts in said bloodaccording to step a) with the level of each of said one or more genetranscripts in blood from one or more individuals having said stage ofdiabetes, c) comparing the level of each of said one or more genetranscripts in said blood according to step a) with the level of each ofsaid one or more gene transcripts in blood from one or more individualsnot having said stage of diabetes, d) determining whether the level ofsaid one or more gene transcripts of step a) are characterized asclassifying with the levels of said transcripts in step b) as comparedwith levels of said transcripts in step c), wherein said determinationis indicative of said individual of step a) having said stage ofdiabetes.
 17. The method of any one of claims 12-16, wherein each ofsaid one or more markers identifies a transcript of a gene selected fromthe group consisting of the genes listed in Table 3G.
 18. The method ofany one of claims 1-4 and 12-16, wherein said one or more genetranscripts are transcribed from one or more genes selected from thegroup consisting of: a) non-immune response genes, b) genes expressed bynon blood tissue, and c) genes expressed by non lymphoid tissue.
 19. Themethod of any one of claims 12-16, wherein said diabetes is eithersymptomatic or asymptomatic.
 20. The method of any one of claims 1-4 and12-16, wherein said diabetes is type II diabetes.
 21. The method of anyone of claims 1-4 and 12-16, wherein said blood comprises a blood sampleobtained from said one or more individuals.
 22. The method of claim 21,wherein said blood sample consists of whole blood.
 23. The method ofclaim 21, wherein said blood sample consists of a drop of blood.
 24. Themethod of claim 21, wherein said blood sample consists of blood that hasbeen lysed.
 25. The method of claim 21, further comprising the step ofisolating RNA from said blood samples.
 26. The method of any one ofclaims 1-4 and 12-16, wherein the step of determining the level of eachof said one or more gene transcripts comprises quantitative RT-PCR(QRT-PCR), wherein said one or more transcripts are from step a) and/orstep b) of claims 1-4 and 12-16.
 27. The method of claim 26, whereinsaid QRT-PCR comprises primers which hybridize to said one or moretranscripts or the complement thereof, wherein said one or moretranscripts are from step a) and/or step b) of claims 1-4 and 12-16. 28.The method of claim 27, wherein said primers are 15-25 nucleotides inlength.
 29. The method of claim 27, wherein said primers hybridize toone or more transcripts of one or more genes selected from the group ofgenes listed in Table 3G, or the complement thereof.
 30. The method ofany one of claims 1-4 and 12-16, wherein the step of determining thelevel of each of said one or more gene transcripts comprises hybridizinga first plurality of isolated nucleic acid molecules that correspond tosaid one or more transcripts, to an array comprising a second pluralityof isolated nucleic acid molecules.
 31. The method of claim 30, whereinsaid first plurality of isolated nucleic acid molecules comprises RNA,DNA, cDNA, PCR products or ESTs.
 32. The method of claim 30, whereinsaid array comprises a plurality of isolated nucleic acid moleculescomprising RNA, DNA, cDNA, PCR products or ESTs.
 33. The method of claim32, wherein said array comprises two or more of the markers of claim 1.34. The method of claim 32, wherein said array comprises two or more ofthe markers of claim
 2. 35. The method of claim 32, wherein said arraycomprises two or more of the markers of claim
 3. 36. The method of claim32, wherein said array comprises two or more of the markers of claim 4.37. The method of claim 32, wherein said array comprises a plurality ofnucleic acid molecules that correspond to genes of the human genome. 38.The method of claim 32, wherein said array comprises a plurality ofnucleic acid molecules that correspond to two or more sequences of twoor more genes selected from the group of genes listed in Table 3G.
 39. Aplurality of isolated nucleic acid molecules that correspond to two ormore of the markers of claim
 1. 40. A plurality of isolated nucleic acidmolecules that correspond to two or more of the markers of claim
 2. 41.A plurality of isolated nucleic acid molecules that correspond to two ormore of the markers of claim
 3. 42. A plurality of isolated nucleic acidmolecules that correspond to two or more of the markers of claim
 4. 43.The method of claim 31, wherein said ESTs comprise a length of greaterthan 100 nucleotides.
 44. An array consisting essentially of theplurality of nucleic acid molecules of claim
 39. 45. An array consistingessentially of the plurality of nucleic acid molecules of claim
 40. 46.An array consisting essentially of the plurality of nucleic acidmolecules of claim
 41. 47. An array consisting essentially of theplurality of nucleic acid molecules of claim
 42. 48. A kit fordiagnosing or prognosing diabetes comprising: a) two gene-specificpriming means designed to produce double stranded DNA complementary to agene that corresponds to a marker selected from the group consisting ofmarkers of claim 1, claim 2, claim 3, and claim 4; wherein said firstpriming means contains a sequence which can hybridize to RNA, cDNA or anEST complementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity; c) an enzyme withthermostable DNA polymerase activity; and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject.
 49. A kit for monitoring a course oftherapeutic treatment of diabetes, comprising: a) two gene-specificpriming means designed to produce double stranded DNA complementary to agene that corresponds to a marker selected from the group consisting ofmarkers of claim 1, claim 2, claim 3, and claim 4; wherein said firstpriming means contains a sequence which can hybridize to RNA, cDNA or anEST complementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity; c) an enzyme withthermostable DNA polymerase activity; and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject.
 50. A kit for monitoring progression orregression of diabetes, comprising: a) two gene-specific priming meansdesigned to produce double stranded DNA complementary to a gene thatcorresponds to a marker selected from the group consisting of markers ofclaim 1, claim 2, claim 3, and claim 4; wherein said first priming meanscontains a sequence which can hybridize to RNA, cDNA or an ESTcomplementary to said gene to create an extension product and saidsecond priming means capable of hybridizing to said extension product;b) an enzyme with reverse transcriptase activity; c) an enzyme withthermostable DNA polymerase activity; and d) a labeling means; whereinsaid primers are used to detect the quantitative expression levels ofsaid gene in a test subject.
 51. The kit of any one of claims 48 to 50wherein said gene-specific priming means identified in step a) isselected from the group of genes listed in Table 3G.
 52. A plurality ofnucleic acid molecules that identify or correspond to two or moresequences of two or more genes selected from the group of genes listedin Table 3G.
 53. The method of claim 32, wherein said ESTs comprise alength of greater than 100 nucleotides.